7-acylamino-3-substituted cephalosporanic acid derivatives

ABSTRACT

The compound 7[2-(aminothiazol-4-yl)-2-methyoxyiminoacetamido]-methylthiomethyl-3-cephem-4-carboxylic acid or carboxylic ester thereof, or a pharmaceutically acceptable salt thereof is disclosed as an antimicrobial agent.

This is a division, of application Ser. No. 206,831, filed Nov. 14, 1980now U.S. Pat. No. 4,409,215 which is in turn a continuation, ofapplication Ser. No. 489,236, filed Apr. 28, 1983 now abandoned.

The present invention relates to novel 7-acylamino-3-substitutedcephalosporanic acid derivatives and pharmaceutically acceptable saltsthereof.

More particularly, it relates to novel 7-acylamino-3-substitutedcephalosporanic acid derivatives and pharmaceutically acceptable saltsthereof, which have antimicrobial activity, to processes for thepreparation thereof, to a pharmaceutical composition comprising thesame, and to a method of using the same therapeutically in the treatmentof infectious diseases in human being and animals.

Accordingly, one object of the present invention is to provide novel7-acylamino-3-substituted cephalosporanic acid derivatives andpharmaceutically acceptable salts thereof, which are highly activeagainst a number of pathogenic microorganisms and are useful asantimicrobial agents, especially oral administration.

Another object of the present invention is to provide processes for thepreparation of novel 7-acylamino-3-substituted cephalosporanic acidderivatives and salts thereof.

A further object of the present invention is to provide a pharmaceuticalcomposition comprising, as active ingredients, said7-acylamino-3-substituted cephalosporanic acid derivatives andpharmaceutically acceptable salts thereof.

Still further object of the present invention is to provide a method ofusing said 7-acylamino-3-substituted cephalosporanic acid derivativesand pharmaceutically acceptable salts thereof in the treatment ofinfectious diseases by pathogenic microorganisms in human being andanimals.

The object 7-acylamino-3-substituted cephalosporanic acid derivativesare novel and can be represented by the following general formula:##STR1## in which R¹ is a group of the formula: ##STR2## wherein R⁴ islower alkyl and

R⁵ is amino or a protected amino group,

R² is lower alkoxymethyl, lower alkylthiomethyl or loweralkenylthiomethyl,

R³ is carboxy or a protected carboxy group, and

A is lower alkylene which may have a substituent selected from thegroups consisting of amino, a protected amino group, hydroxy, oxo and agroup of the formula: ═N˜OR⁶, wherein R⁶ is hydrogen, lower alkenyl,lower alkynyl, lower alkyl or lower alkyl substituted by one or moresubstituent(s) selected from carboxy, a protected carboxy group, amino,a protected amino group, and heterocyclic group.

In the object compounds (I) and the corresponding starting compounds(II) to (IV) in Processes 1 and 7 mentioned below, it is to beunderstood that there may be one or more stereoisomeric pair(s) such asoptical and geometrical isomers due to asymmetric carbon atom and doublebond in those molecules and such isomers are also included within thescope of the present invention.

With regard to geometrical isomers in the object compounds and thestarting compounds, it is to be noted that, for example, the objectcompounds, wherein A means a group of the formula: ═C═N˜OR⁶, include synisomer, anti isomer and a mixture thereof, and the syn isomer means onegeometrical isomer having the partial structure represented by thefollowing formula: ##STR3## wherein R¹ and R⁶ are each as defined above,and the anti isomer means the other geometrical isomer having thepartial structure represented by the following formula: ##STR4## whereinR¹ and R⁶ are each as defined above.

Regarding the other object and starting compounds as mentioned above,the syn isomer and the anti isomer can also be referred to the samegeometrical isomers as illustrated for the compounds (I).

Suitable pharmaceutically acceptable salts of the object compounds (I)are conventional non-toxic salts and may include a salt with a base oran acid addition salt such as a salt with an inorganic base, forexample, an alkali metal salt (e.g. sodium salt, potassium salt, etc.),an alkaline earth metal salt (e.g. calcium salt, magnesium salt, etc.),an ammonium salt; a salt with an organic base, for example, an organicamine salt (e.g. triethylamine salt, pyridine salt, picoline salt,ethanolamine salt, triethanolamine salt, dicyclohexylamine salt,N,N'-dibenzylethylenediamine salt, etc.) etc.; an inorganic acidaddition salt (e.g. hydrochloride, hydrobromide, sulfate, phosphate,etc.); an organic carboxylic or sulfonic acid addition salt (e.g.formate, acetate, trifluoroacetate, maleate, tartrate, methanesulfonate,benzenesulfonate, p-toluenesulfonate, etc.); a salt with a basic oracidic amino acid (e.g. arginine, aspartic acid, glutamic acid, etc.);an intermolecular or intramolecular quaternary salt, and the like. Thesaid intermolecular quaternary salt can be formed in case that theheterocyclic group in R⁶ in the compounds (I) contains nitrogen atom(s)(e.g. pyridyl, etc.), and suitable intermolecular quaternary salt mayinclude 1-lower alkylpyridinium lower alkylsulfate (e.g.1-methylpyridinium methylsulfate, 1-ethylpyridinium ethylsulfate, etc.),1-lower alkylpyridinium halide (e.g. 1-methylpyridinium iodide, etc.)and the like. The said intramolecular salt can be formed in case thatheterocyclic group in R⁶ in the compounds (I) contains nitrogen atom(s)(e.g. pyridyl etc.) and R³ is carboxy, and suitable intramolecular saltmay include 1-lower alkylpyridinium carboxylate (e.g. 1-methylpyridiniumcarboxylate, 1-ethylpyridinium carboxylate, 1-propylpyridiniumcarboxylate, 1-isopropylpyridinium carboxylate, 1-butylpyridiniumcarboxylate, etc.); and the like.

According to the present invention, the object compounds (I) and thepharmaceutically acceptable salts thereof can be prepared by theprocesses as illustrated by the following reaction schemes. ##STR5## inwhich R¹, R², R³, R⁵ and A are each as defined above,

R_(a) ¹ is a group of the formula: ##STR6## wherein R⁸ is a protectedamino group, R_(b) ¹ is a group of the formula: ##STR7## R_(a) ³ is aprotected carboxy group, R_(b) ³ is lower alkoxycarbonyl substituted byprotected amino and protected carboxy groups,

R_(c) ³ is lower alkoxycarbonyl substituted by amino and carboxy,

R⁷ is lower alkyl,

A¹ is lower alkylene having a protected amino group,

A² is lower alkylene having an amino group,

A³ is lower alkylene having an oxo group,

A⁴ is lower alkylene having a hydroxy group,

A⁵ is lower alkylene which may have a group of the formula ═N˜OR⁶,wherein R⁶ is as defined above,

A⁶ is lower alkylene having a group of the formula: ═N˜OR_(a) ⁶, whereinR_(a) ⁶ is lower alkyl substituted by a protected carboxy group,

A⁷ is lower alkylene having a group of the formula: ═N˜OR_(b) ⁶, whereinR_(b) ⁶ is lower alkyl substituted by carboxy,

A⁸ is lower alkylene having a group of the formula: ═N˜OR_(c) ⁶, whereinR_(c) ⁶ is lower alkoxycarbonyl(lower)alkyl substituted by protectedamino and protected carboxy groups or lower alkyl substituted byprotected amino and protected carboxy groups,

A⁹ is lower alkylene having a group of the formula: ═N˜OR_(d) ⁶, whereinR_(d) ⁶ is lower alkoxycarbonyl(lower)alkyl substituted by amino andcarboxy or lower alkyl substituted by amino and carboxy,

A¹⁰ is lower alkylene having a group of the formula: ═N˜OR_(e) ⁶,wherein R_(e) ⁶ is lower alkyl substituted by a group of the formula:##STR8## A¹¹ is lower alkylene having a group of the formula: ═N˜OR_(f)⁶, wherein R_(f) ⁶ is lower alkyl substituted by a group of the formula:##STR9## wherein R⁷ is as defined above, A¹² is lower alkylene having agroup of the formula: ═N˜OR_(g) ⁶, wherein R_(g) ⁶ is lower alkylsubstituted by a cation of the formula: ##STR10## wherein R⁷ is asdefined above, and X¹ is halogen.

Some of the starting compounds (II), (III) and (IV) used in Processes 1and 7 are new and can be represented by the following formula: ##STR11##in which R^(A) is amino or a protected amino group,

R^(b') is lower alkenyl, X is --S-- or --SO--, and

R³ is as defined above, and a salt thereof.

Suitable salts of the above starting compound are the same as thoseexemplified for the object compounds (I).

The starting compound thus formulated and other starting compounds canbe prepared, for example, from the known compounds by the methods in thefollowing reaction schemes, and others can be prepared in a similarmanner thereto or in a conventional manner. ##STR12## in which R_(a) ¹,R_(b) ¹, R², R³, R_(b) ³, R⁶, A¹, A², A³, A⁵ and X¹ are each as definedabove,

R^(a) is a protected amino group,

R^(b) is lower alkyl or lower alkenyl,

R^(c) is a protected carboxy group or carboxy,

R^(d) is trihalomethyl,

Y is a conventional group which is capable to be replaced by the residue(--SR^(b)) of the compound of the formula: HS--R^(b), in which R^(b) isas defined above,

A¹³ is lower alkylene having a group of the formula: ═N˜OR⁶, wherein R⁶is as defined above, and

R_(h) ⁶ is lower alkyl substituted by a protected carboxy group, loweralkoxycarbonyl(lower)alkyl substituted by protected amino and protectedcarboxy groups or lower alkyl substituted by protected amino andprotected carboxy groups.

In the above and subsequent description of the present specification,suitable examples and illustration of the various definitions to beincluded within the scope thereof are explained in detail as follows.

The term "lower" in the present specification is intended to mean agroup having 1 to 6 carbon atoms, unless otherwise indicated.

Suitable "lower alkyl" group may include straight or branched one suchas methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, isopentyl,neopentyl, hexyl and the like, in which the preferred one is C₁ -C₃alkyl.

Suitable "lower alkenyl" group may include straight or branched one suchas vinyl, 1-propenyl, allyl, 1-(or 2- or 3-)butenyl, 1-(or 2- or 3- or4-)pentenyl, 1-(or 2- or 3- or 4- or 5-)hexenyl, 2-methyl-2-propenyl,and the like, in which the preferred one is C₂ -C₅ alkenyl.

Suitable "lower alkynyl" group may include straight or branched one suchas propargyl, 2-(or 3-)butynyl, 2-(or 3- or 4-)pentynyl, 2-(or 3- or 4-or 5-)hexynyl, and the like, in which the preferred one is C₂ -C₅alkynyl.

Suitable "lower alkoxymethyl" group may include a methyl groupsubstituted by straight or branched lower alkoxy such as methoxymethyl,ethoxymethyl, propoxymethyl, isopropoxymethyl, pentyloxymethyl,hexyloxymethyl, and the like, in which the preferred one is C₁ -C₃alkoxymethyl.

Suitable "lower alkylthiomethyl" group may include a methyl groupsubstituted by straight or branched lower alkylthio such asmethylthiomethyl, ethylthiomethyl, propylthiomethyl, isobutylthiomethyl,pentylthiomethyl, hexlthiomethyl, and the like, in which the preferredone is C_(1-C) ₃ alkylthiomethyl.

Suitable "lower alkenylthiomethyl" group may include a methyl groupsubstituted by straight or branched lower alkenylthio such asvinylthiomethyl, 1-propenylthiomethyl, allylthiomethyl,2-methyl-2-propenylthiomethyl, 1-(or 2- or 3-)butenylthiomethyl, 1-(or2- or 3- or 4-)pentenylthiomethyl, 1-(or 2- or 3- or4-)hexenylthiomethyl, and the like, in which the preferred one is C₂ -C₅alkenylthiomethyl.

Suitable "protected amino group" may include an amino group substitutedby a conventional amino-protective group which is used in penicillin andcephalosporin compounds, for example, acyl as mentioned below, mono(ordi or tri)phenyl(lower)alkyl (e.g. benzyl, benzhydryl, trityl, etc.),lower alkoxycarbonyl(lower)alkylidene or its enamine tautomer (e.g.1-methoxycarbonyl-1-propen-2-yl, etc.), di(lower)alkylaminomethylene(e.g. dimethylaminomethylene, etc.), etc.

Suitable "acyl" may include an aliphatic acyl, an aromatic acyl, aheterocyclic acyl and an aliphatic acyl substituted with aromatic orheterocyclic group(s).

The aliphatic acyl may include saturated or unsaturated, acyclic orcyclic ones, such as lower alkanoyl (e.g. formyl, acetyl, propionyl,butyryl, isobutyryl, valeryl, isovaleryl, pivaloyl, hexanoyl, etc.),lower alkanesulfonyl (e.g. mesyl, ethanesulfonyl, propanesulfonyl,etc.), lower alkoxycarbonyl (e.g. methoxycarbonyl, ethoxycarbonyl,propoxycarbonyl, butoxycarbonyl, tert-butoxycarbonyl, etc.), loweralkenoyl (e.g. acryloyl, methacryloyl, crotonoyl, etc.), (C₃-C₇)-cycloalkanecarbonyl (e.g. cyclohexanecarbonyl, etc.), amidino, andthe like.

The aromatic acyl may include aroyl (e.g. benzoyl, toluoyl, xyloyl,etc.), arenesulfonyl (e.g. benzenesulfonyl, tosyl, etc.), and the like.

The heterocyclic acyl may include heterocyclecarbonyl (e.g. furoyl,thenoyl, nicotinoyl, isonicotinoyl, thiazolycarbonyl,thiadiazolycarbonyl, tetrazolylcarbonyl, etc.), and the like.

The aliphatic acyl substituted with aromatic group(s) may includephenyl(lower)alkanoyl (e.g. phenylacetyl, phenylpropionyl,phenylhexanolyl, etc.), phenyl(lower)alkoxycarbonyl (e.g.benzyloxycarbonyl, phenethyloxycarbonyl, etc.), phenoxy(lower)alkanoyl(e.g. phenoxyacetyl, phenoxypropionyl, etc.), and the like.

The aliphatic acyl substituted with heterocyclic group(s) may includethienylacetyl, imidazolylacetyl, furylacetyl, tetrazolylacetyl,thiazolylacetyl, thiadiazolylacetyl, thienylpropionyl,thiadiazolylpropionyl, and the like.

These acyl groups may be further substituted with one or more suitablesubstituents such as lower alkyl (e.g. methyl, ethyl, propyl, isopropyl,butyl, pentyl, hexyl, etc.), halogen (e.g. chlorine, bromine, iodine,fluorine), lower alkoxy (e.g. methoxy, ethoxy, propoxy, isipropoxy,butoxy, pentyloxy, hexyloxy, etc.), lower alkylthio (e.g. methylthio,ethylthio, propylthio, isopropylthio, butylthio, pentylthio, hexylthio,etc.), nitro and the like, and preferable acyl having suchsubstituent(s) may be mono(or di or tri)halo(lower)alkanoyl (e.g.chloroacetyl, bromoacetyl, dichloroacetyl, trifluoroacetyl, etc.),mono(or di or tri)halo(lower)alkoxycarbonyl (e.g. chloromethoxycarbonyl,dichloromethoxycarbonyl, 2,2,2-tri-chloroethoxycarbonyl, etc.), nitro(or halo or lower alkoxy)phenyl(lower)alkoxycarbonyl (e.g.nitrobenzyloxycarbonyl, chlorobenzyloxycarbonyl,methoxybenzyloxycarbonyl, etc.), and the like.

Suitable "protected carboxy group" may include an esterified carboxygroup which is conventionally used in penicillin or cephalosporincompounds at their 3rd or 4th position thereof.

Suitable "ester moiety" in "esterified carboxy group" may include loweralkyl ester (e.g. methyl ester, ethyl ester, propyl ester, isopropylester, butyl ester, isobutyl ester, t-butyl ester, pentyl ester,tertpentyl ester, hexyl ester, etc.), lower alkenyl ester (e.g. vinylester, allyl ester, etc.), lower alkynyl ester (e.g. ethynyl ester,propynyl ester, etc.), lower alkoxy(lower)alkyl ester (e.g.methoxymethyl ester, ethoxymethyl ester, isopropoxymethyl ester,1-methoxyethyl ester, 1-ethoxyethyl ester, etc.), loweralkylthio(lower)alkyl ester (e.g. methylthiomethyl ester,ethylthiomethyl ester, ethylthioethyl ester isopropylthiomethyl ester,etc.), amino- and carboxy substituted lower alkyl ester (e.g.2-amino-2-carboxyethyl ester, 3-amino-3-carboxypropyl ester, etc.),protected amino and protected carboxy substituted lower alkyl ester suchas lower alkoxycarbonylamino and mono(or di ortri)phenyl(lower)alkoxycarbonyl substituted lower alkyl ester (e.g.2-tert-butoxycarbonylamino-2-benzhydryloxycarbonylethyl,3-tert-butoxycarbonylamino-3-benzhydryloxycarbonylpropyl, etc.), mono(ordi or tri)halo(lower)alkyl ester (e.g. 2-iodoethyl ester,2,2,2-trichloroethyl ester, etc.), lower alkanoyloxy(lower)alkyl ester(e.g. acetoxymethyl ester, propionyloxymethyl ester, butyryloxymethylester, isobutyryloxymethyl ester, valeryloxymethyl ester,pivaloyloxymethyl ester, hexanoyloxymethyl ester, 2-acetoxyethyl ester,2-propionyloxyethyl ester, 1-acetoxypropyl ester, etc.), loweralkanesulfonyl(lower)alkyl ester (e.g. mesylmethyl ester, 2-mesylethylester, etc.), mono(or di or tri)phenyl(lower)alkyl ester which may haveone or more suitable substituent(s) (e.g. benzyl ester, 4-methoxybenzylester, 4-nitrobenzyl ester, phenethyl ester, benzhydryl ester, tritylester, bis(methoxyphenyl)methyl ester, 3,4-dimethoxybenzyl ester,4-hydroxy-3,5-di-t-butylbenzyl ester, etc.), aryl ester which may haveone or more suitable substituents (e.g. phenyl ester, tolyl ester,t-butylphenyl ester, xylyl ester, mesityl ester, cumenyl ester, salicylester, etc.), heterocyclic ester (e.g. phthalidyl ester, etc.) and thelike.

Suitable "lower alkylene" group may include straight or branched onesuch as methylene, ethylene, trimethylene, propylene, tetramethylene,hexamethylene, and the like, in which the preferred one is C₁ -C₂alkylene and the most preferred one is methylene.

Suitable "heterocyclic" group may include saturated or unsaturated,monocyclic or polycyclic heterocyclic group containing at least onehetero-atom such as an oxygen, sulfur, nitrogen atom and the like. And,especially preferable heterocyclic group may be heterocyclic group suchas

unsaturated 3 to 8-membered (more preferably 5 or 6-membered)heteromonocyclic group containing 1 to 4 nitrogen atom(s), for example,pyrrolyl, pyrrolinyl, imidazolyl, pyrazolyl, pyridyl and its N-oxide,dihydropyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazolyl (e.g.4H-1,2,4-triazolyl, 1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, etc.),tetrazolyl (e.g. 1H-tetrazolyl, 2H-tetrazolyl, etc.) etc.;

saturated 3 to 8-membered (more preferably 5 or 6-membered)heteromonocyclic group containing 1 to 4 nitrogen atom(s), for example,pyrrolidinyl, imidazolidinyl, piperidino, piperazinyl, etc.:

unsaturated condensed heterocyclic group containing 1 to 4 nitrogenatom(s), for example, indolyl, isoindolyl, indolizinyl, benzimidazolyl,quinolyl, isoquinolyl, indazolyl, benzotriazolyl, etc.;

unsaturated 3 to 8-membered (more preferably 5 or 6-membered)heteromonocyclic group containing 1 to 2 oxygen atom(s) and 1 to 3nitrogen atom(s), for example, oxazolyl, isoxazolyl, oxadiazolyl (e.g.1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,5-oxadiazolyl, etc.), etc.;

saturated 3 to 8-membered (more preferably 5 or 6-membered)heteromonocyclic group containing 1 to 2 oxygen atom(s) and 1 to 3nitrogen atom(s), for example, morpholinyl, sydnonyl, etc.;

unsaturated condensed heterocycyclic group containing 1 to 2 oxygenatom(s) and 1 to 3 nitrogen atom(s), for example, benzoxazolyl,benzoxadiazolyl, etc.;

unsaturated 3 to 8-membered (more preferably 5 or 6-membered)heteromonocyclic group containing 1 to 2 sulfur atom(s) and 1 to 3nitrogen atom(s), for example, thiazolyl, isothiazolyl, thiadiazolyl(e.g. 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl,1,2,5-thiadiazolyl, etc.), dihydrothiazinyl, etc.

saturated 3 to 8-membered (more preferably 5 or 6-membered)heteromonocyclic group containing 1 to 2 sulfur atom(s) and 1 to 3nitrogen atom(s), for example, thiazolidinyl, etc.;

unsaturated 3 to 8-membered (more preferably 5 or 6-membered)heteromonocyclic group containing 1 to 2 sulfur atom(s), for example,thienyl, dihydrodithiinyl, etc.,;

unsaturated condensed heterocyclic group containing 1 to 2 sulfuratom(s) and 1 to 3 nitrogen atom(s), for example, benzothiazolyl,benzothiadiazolyl, etc.;

unsaturated 3 to 8-membered (more preferably 5 or 6-membered)heteromonocyclic group containing an oxygen atom, for example, furyl,etc.;

unsaturated 3 to 8-membered (more preferably 5 or 6-membered)heteromonocyclic group containing an oxygen atom and 1 to 2 sulfuratom(s), for example, dihydrooxathiinyl, etc.;

unsaturated condensed heterocyclic group containing 1 to 2 sulfuratom(s), for example, benzothienyl, benzodithiinyl, etc.;

unsaturated condensed heterocyclic group containing an oxygen atom and 1to 2 sulfur atom(s), for example, benzoxathiinyl, etc. and the like.

Thus defined heterocyclic group may optionally be substituted by one toten, same or different, suitable substituent(s) such as: lower alkyl(e.g. methyl, ethyl, etc.); lower alkoxy (e.g. methoxy, ethoxy, propoxy,etc.); lower alkylthio (e.g. methylthio, ethylthio, etc.); loweralkylamino (e.g. methylamino, etc.); cyclo(lower)alkyl (e.g.cyclopentyl, cyclohexyl, etc.); cyclo(lower)alkenyl (e.g. cyclohexenyl;cyclohexadienyl, etc.); hydroxy; halogen (e.g. chloro, bromo, etc.);amino; protected amino as aforementioned; cyano; nitro; carboxy;protected carboxy as aforementioned; sulfo; sulfamoyl; imino; oxo;amino(lower)alkyl (e.g. aminomethyl, aminoethyl, etc.); and the like.

Suitable "lower alkoxycarbonyl(lower)alkyl" group may includeethoxycarbonylmethyl, propoxycarbonylmethyl, 1- or2-ethoxycarbonylethyl, and the like.

Suitable "lower alkoxycarbonyl" moiety may include ethoxycarbonyl,propoxycarbonyl, and the like.

Suitable "halogen" may include chloro, bromo, iodo, and the like.

Suitable "conventional group which is capable to be replaced by theresidue (--S--R^(b)) of the compound of the formula: HS--R^(b) " mayinclude halogen as exemplified above.

Suitable "trihalomethyl" may include trichloromethyl, and the like.

Particularly, the preferred embodiment of the symbols "R¹ --A--", "R² "and "R³ " of the object compounds (I) can be represented as follows.

The symbol "R¹ --A--" can be represented by the formulae:

○1 ##STR13## in which R¹ is a group of the formula: ##STR14## wherein R⁵is amino or acylamino [more preferably lower alkanamido (e.g. formamido,acetamido, propionamido, etc.) or mono-or di- ortrihalo(lower)alkanamido (e.g. chloroacetamido, dichloroacetamido,trifluoroacetamido, etc.)],

R⁶ is lower alkyl (e.g. methyl, ethyl, propyl, isopropyl, butyl, pentyl,hexyl, etc.), carboxy(lower)alkyl (e.g. carboxymethyl, 1- or2-carboxyethyl, 1- or 2- or 3- carboxypropyl, etc.), or esterifiedcarboxy(lower)alkyl [more preferably lower alkoxycarbonyl(lower)alkyl(e.g. methoxycarbonylmethyl, ethoxycarbonylmethyl, tert-butoxycarbonylmethyl, tert- butoxycarbonylethyl, etc.) or mono- or di-or triphenyl(lower)alkoxycarbonyl(lower)alkyl (e.g.benzyloxycarbonylmethyl, benzhydryloxycarbonylmethyl,benzhydryloxycarbonylethyl etc.)]; or

○2 R¹ --A--, in which R¹ is a group of the formula: ##STR15## wherein R⁵is amino or acylamino [more preferably lower alkanamido (e.g. formamido,acetamido, propionamido, etc.)], and

A is methylene, aminomethylene, acylaminomethylene [more preferablylower alkoxycarbonylaminomethylene (e.g. methoxycarbonylaminomethylene,ethoxycarbonylaminomethylene, tert- butoxycarbonylaminomethylene,etc.)], hydroxymethylene or carbonyl; or

○3 R¹ --A--, in which R¹ is a group of the formula: ##STR16## wherein R⁴is lower alkyl (e.g. methyl, ethyl, propyl, isopropyl, butyl, pentyl,etc.), and

A is aminomethylene or acylaminomethylene [more preferably loweralkoxycarbonylaminomethylene (e.g. methoxycarbonylaminomethylene,ethoxycarbonylaminomethylene, tert- butoxycarbonylaminomethylene,etc.)].

The symbol "R² " can be represented by: lower alkoxymethyl (e.g.methoxymethyl, ethoxymethyl, propoxymethyl, isopropoxymethyl, etc.);lower alkylthiomethyl (e.g. methylthiomethyl, ethylthiomethyl,propylthiomethyl, isopropylthiomethyl, etc.); or lower alkenylthiomethyl(e.g. vinylthiomethyl, allylthiomethyl, butenylthiomethyl, etc.).

The symbol "R³ " can be represented by: carboxy or esterified carboxy[more preferably lower alkoxycarbonyl (e.g. methoxycarbonyl,ethoxycarbonyl, tert-butoxycarbonyl, etc.) or mono- or di- ortriphenyl(lower)alkoxycarbonyl (e.g. benzyloxycarbonyl,benzhydryloxycarbonyl, phenethyloxycarbonyl, etc.)].

The processes 1 to 13 for the preparation of the object compounds (I) ofthe present invention are explained in detail in the following.

(1) Process 1:

The compounds (I) or a salt thereof can be prepared by reacting thecompound (II) or its reactive derivative at the amino group or a saltthereof with the compound (III) or its reactive derivative at thecarboxy group or a salt thereof.

Suitable salts of the starting compounds (II) and (III) may include thesame ones as illustrated for the compounds (I).

Suitable reactive derivative at the carboxy group of the compound (II)may include a conventional one, for example, a silyl derivative formedby the reaction of the compound (II) with a silyl compound such asbis(trimethylsilyl)acetamide, trimethylsilylacetamide, etc.; isocyanate;isothiocyanate; Schiff's base or its tautomeric enamine type isomerformed by the reaction of the amino group with a carbonyl compound suchas an aldehyde compound (e.g. acetaldehyde, isopentaldehyde,benzaldehyde, salicylaldehyde, phenylacetaldehyde, p-nitrobenzaldehyde,m-chlorobenzaldehyde, p-chlorobenzaldehyde, hydroxynaphthoaldehyde,furfural, thiophenecarboaldehyde, etc.) or a ketone compound (e.g.acetone, methyl ethyl ketone, methyl isobutyl ketone, acetylacetone,ethyl acetoacetate, etc.), and the like.

Suitable reactive derivative of the compound (III) may include, forexample, an acid halide, an acid anhydride, an activated amide, anactivated ester, and the like, and preferably an acid chloride and acidbromide; a mixed acid anhydride with an acid such as substitutedphosphoric acid (e.g. dialkylphosphoric acid, phenylphosphoric acid,diphenylphosphoric acid, dibenzylphosphoric acid, halogenated phosphoricacid, etc.), dialkylphosphorous acid, sulfurous acid, thiosulfuric acid,sulfuric acid, alkyl carbonate (e.g. methyl carbonate, ethyl carbonate,propyl carbonate, etc.), aliphatic carboxylic acid (e.g. pivalic acid,pentanoic acid, isopentanoic acid, 2-ethylbutyric acid, trichloroaceticacid, etc.), aromatic carboxylic acid (e.g. benzoic acid, etc.); asymmetrical acid anhydride; an activated acid amide with a heterocycliccompound containing imino function such as imidazole, 4-substitutedimidazole, dimethylpyrazole, triazole or tetrazole; an activated ester(e.g. p-nitrophenyl ester, 2,4-dinitrophenyl ester, trichlorophenylester, pentachlorophenyl ester, mesylphenyl ester, phenylazophenylester, phenyl thioester, p-nitrophenyl thioester, p-cresyl thioester,carboxymethyl thioester, pyridyl ester, piperidinyl ester, 8-quinolylthioester, or an ester with a N-hydroxy compound such asN,N-dimethylhydroxylamine, 1-hydroxy-2-(1H)-pyridone,N-hydroxysuccinimide, N-hydroxyphthalimide, 1-hydroxybenzotriazole,1-hydroxy-6-chlorobenzotriazole, etc.), and the like.

Additionally, as a reactive derivative of the compound (III), wherein Ais aminomethylene, the compound of the following formula can also beused. ##STR17##

The suitable reactive derivative can optionally be selected from theabove according to the kinds of the compounds (II) and (III) to be usedpractically.

This reaction can be carried out in the presence of an organic orinorganic base such as alkali metal (e.g. lithium, sodium, potassium,etc.), alkaline earth metal (e.g. calcium, etc.), alkali metal hydride(e.g. sodium hydride, etc.), alkaline earth metal hydride (e.g. calciumhydride, etc.), alkaline metal hydroxide (e.g. sodium hydroxide,potassium hydroxide, etc.), alkali metal carbonate (e.g. sodiumcarbonate, potassium carbonate, etc.), alkali metal bicarbonate (e.g.sodium bicarbonate, potassium bicarbonate, etc.), alkali metal alkoxide(e.g. sodium methoxide, sodium ethoxide, potassium tert-butoxide, etc.),trialkylamine (e.g. triethylamine, etc.), pyridine compound (e.g.pyridine, lutidine, picoline, etc.), quinoline, and the like.

In case that the compound (III) is used in a form of the free acid or asalt in this reaction, the reaction is preferably carried out in thepresence of a condensing agent such as a carbodiimide compound [e.g.N,N'-dicyclohexylcarbodiimide,N-cyclohexyl-N'-morpholinoethylcarbodiimide,N-cyclohexyl-N'-(4-diethylaminocyclohexyl)carbodiimide,N,N'-diethylcarbodiimide, N,N'-diisopropylcarbodiimide,N-ethyl-N'-(3-dimethylaminopropyl)carbodiimide, etc.], a keteniminecompound (e.g. N,N'-carbonylbis(2-methylimidazole),pentamethyleneketene-N-cyclohexylimine,diphenylketene-N-cyclohexylimine, etc.); an olefinic or acetylenic ethercompounds (e.g. ethoxyacetylene, β-chlorovinylethyl ether), a sulfonicacid ester of N-hydroxybenzotriazole derivative [e.g.1-(4-chlorobenzenesulfonyloxy)-6-chloro-1H-benzotriazole, etc.], acombination of trialkylphosphite or triphenylphosphine and carbontetrachloride, disulfide or diazenedicarboxylate (e.g. diethyldiazenedicarboxylate, etc.), a phosphorus compound (e.g. ethylpolyphosphate, isopropyl polyphosphate, phosphoryl chloride, phosphorustrichloride, etc.), thionyl chloride, oxalyl chloride,N-ethylbenzisoxazolium salt, N-ethyl-5-phenylisoxazolium-3-sulfonate, areagent (referred to as so-called "Vilsmeier reagent") formed by thereaction of an amide compound such as dimethylformamide,N-methylformamide or the like with a halogen compound such as thionylchloride, phosphoryl chloride, phosgene or the like.

The reaction is usually carried out in a conventional solvent which doesnot adversely influence the reaction such as water, acetone, dioxane,acetonitrile, chloroform, benzene, methylene chloride, ethylenechloride, tetrahydrofuran, ethyl acetate, N,N-dimethylformamide,pyridine, hexamethylphosphoramide, etc., or a mixture thereof.

Among these solvents, hydrophilic solvents may be used in a mixture withwater.

The reaction temperature is not critical and the reaction is usuallycarried out under cooling to warming.

(2) Process 2:

The compound (I-b) or a salt thereof can be prepared by subjecting thecompound (I-a) or a salt thereof to removal reaction of theamino-protective group in A¹.

Suitable method for this removal reaction may include conventional onesuch as hydrolysis, reduction and the like.

(i) For Hydrolysis:

Hydrolysis is preferably carried out in the presence of an acid.

Suitable acid may be an inorganic acid (e.g. hydrochloric acid,hydrobromic acid, sulfuric acid, etc.), an organic acid (e.g. formicacid, acetic acid, trifluoroacetic acid, propionic acid, benzenesulfonicacid, p-toluenesulfonic acid, etc.), an acidic ion-exchange resin andthe like. In case that trifluoroacetic acid is used in this reaction,the reaction is preferably carried out in the presence of cationtrapping agents (e.g. anisole, etc.).

The acid suitable for this hydrolysis can be selected according to thekinds of the protective group to be removed, for example, thishydrolysis can preferably be applied to the amino-protective group forA¹ such as substituted or unsubstituted lower alkoxycarbonyl,substituted or unsubstituted lower alkanoyl.

The hydrolysis is usually carried out in a conventional solvent whichdoes not adversely influence the reaction such as water, methanol,ethanol, propanol, tetrahydrofuran, N,N-dimethylformamide, dioxane or amixture thereof, and further the above-mentioned acids can also be usedas a solvent when they are in liquid.

The reaction temperature of this hydrolysis is not critical, and thereaction is usually carried out under cooling to at somewhat elevatedtemperature.

(ii) For Reduction:

Reduction is carried out in a conventional manner, including chemicalreduction and catalytic reduction.

Suitable reducing agents to be used in chemical reduction are acombination of a metal (e.g. tin, zinc, iron, etc.) or metallic compound(e.g. chromium chloride, chromium acetate, etc.) and an organic orinorganic acid (e.g. formic acid, acetic acid, propionic acid,trifluoroacetic acid, p-toluenesulfonic acid, hydrochloric acid,hydrobromic acid, etc.).

Suitable catalysts to be used in catalytic reduction are conventionalones such as platinum catalysts (e.g. platinum plate, spongy platinum,platinum black, colloidal platinum, platinum oxide, platinum wire,etc.), palladium catalysts (e.g. spongy palladium, palladium black,palladium oxide, palladium on carbon, colloidal palladium, palladium onbarium sulfate, palladium on barium carbonate, etc.), nickel catalysts(e.g. reduced nickel, nickel oxide, Raney nickel, etc.), cobaltcatalysts (e.g. reduced cobalt, Raney cobalt, etc.), iron catalysts(e.g. reduced iron, Raney iron, etc.), copper catalysts (e.g. reducedcopper, Raney copper, Ullman copper, etc.) and the like.

The reduction manner can be selected according to the kinds of theprotective group to be removed, for example, the chemical reduction canpreferably be applied to the amino-protective group for A¹ such ashalo(lower)alkoxycarbonyl and the like, and catalytic reduction canpreferably be applied to that such as substituted or unsubstitutedar(lower)alkoxycarbonyl, and the like.

The reduction is usually carried out in a conventional solvent whichdoes not adversely influence the reaction such as water, methanol,ethanol, propanol, N,N-dimethylformamide, or a mixture thereof.Additionally, in case that the above-mentioned acids to be used inchemical reduction are in liquid, they can also be used as a solvent.Further, a suitable solvent to be used in catalytic reduction may be theabove-mentioned solvent, and other conventional solvent such as diethylether, dioxane, tetrahydrofuran, etc., or a mixture thereof.

The reaction temperature of this reduction is not critical and thereaction is usually carried out under cooling to warming.

The present invention includes, within the scope of the invention, casesthat the protected amino group in R¹ and/or the protected carboxy groupfor R³ are transformed into free amino group and/or free carboxy group,respectively during the reaction.

(3) Process 3:

The compound (I-d) or a salt thereof can be prepared by subjecting thecompound (I-c) or a salt thereof to removal reaction of theamino-protective group in R_(a) ¹.

This reaction is carried out by a conventional method such ashydrolysis, reduction, and the like.

The method of hydrolysis and reduction, and the reaction conditions(e.g. reaction temperature, solvent, etc.) are substantially the same asthose illustrated for the removal reaction of the amino-protective groupof the compound (I-a) in Process 2, and therefore are to be referred tosaid explanation.

The present invention includes, within the scope of the invention, casesthat the protected amino group in A and/or the protected carboxygroup(s) for R³ and A are transformed into free amino group and/or freecarboxy group, respectively during the reaction.

(4) Process 4:

The compound (I-f) or a salt thereof can be prepared by subjecting thecompound (I-e) or a salt thereof to removal reaction of thecarboxy-protective group for R_(a) ³.

This reaction is carried out by a conventional method such ashydrolysis, reduction, and the like.

The method of hydrolysis and reduction, and the reaction conditions(e.g. reaction temperature, solvent, etc.) are substantially the same asthose illustrated for the removal reaction of the amino-protective groupof the compound (I-a) in Process 2, and therefore are to be referred tosaid explanation.

The present invention includes, within the scope of the invention, casesthat the protected amino group(s) in R¹ and A and/or the protectedcarboxy group in A are transformed into free amino group(s) and/or afree carboxy group, respectively during the reaction.

(5) Process 5:

The compound (I-e) or a salt thereof can be prepared by introducing acarboxy-protective group into the compound (I-f) or a salt thereof.

The introducing agent of a carboxy-protective group to be used in thisreaction may include a conventional esterifying agent such as an alcoholor its reactive equivalent (e.g. halide, sulfonate, sulfate, diazocompound, etc.), and the like.

This reaction can also be carried out in the presence of a base, andsuitable examples thereof are the same as those given in the explanationof Process 1, and can preferably be carried out in the presence of metaliodide (e.g. sodium iodide, etc.).

This reaction is usually carried out in a conventional solvent whichdoes not adversely influence the reaction such as N,N-dimethylformamide,tetrahydrofuran, dioxane, methanol, ethanol, etc., or a mixture thereof.

The reaction temperature is not critical, and the reaction is usuallycarried out under cooling to at somewhat elevated temperature.

In case that the alcohol is used as the introducing agent of acarboxy-protective group, the reaction can be carried out in thepresence of a condensing agent as illustrated in Process 1.

(6) Process 6:

The compound (I-h) or a salt thereof can be prepared by reducing thecompound (I-g) or a salt thereof.

The reduction can be carried out by a conventional method such asreduction using a reducing agent, catalytic reduction, and the like.

Suitable reducing agent may include a conventional one used forconversion of a carbonyl group to a hydroxymethyl group such as metalborohydride, for example, alkali borohydride (e.g. sodium borohydride,potassium borohydride, sodium cyanoborohydride, etc.), lithium aluminumhydride, etc.; diborane; and the like.

The catalyst to be used in the catalytic reduction may include the sameones as exemplified for the reduction in Process 2.

This reaction is usually carried out in a conventional solvent whichdoes not adversely influence the reaction such as water, methanol,ethanol, tetrahydrofuran, dioxane; etc., or a mixture thereof.

The reaction temperature is not critical and the reaction is usuallycarried out under cooling to warming.

(7) Process 7:

The compound (I-i) or a salt thereof can be prepared by reacting thecompound (IV) or a salt thereof with the compound (V).

Suitable salts of the starting compound (IV) may include the same saltswith a base for the compounds (I).

This reaction is usually carried out in a conventional solvent whichdoes not adversely influence the reaction such as ethyl acetate,methylene chloride, chloroform, carbon tetrachloride, tetrahydrofuran,

The reaction temperature is not critical and the reaction is usuallycarried out under cooling to warming.

(8) Process 8:

The compound (I-l) or a salt thereof can be prepared by subjecting thecompound (I-k) or a salt thereof to removal reaction of thecarboxy-protective group in A⁶.

This reaction is carried out by a conventional method such ashydrolysis, reduction, and the like.

The method of hydrolysis and reduction, and the reaction conditions(e.g. reaction temperature, solvent, etc.) are substantially the same asthose illustrated for the removal reaction of the amino-protective groupof the compound (I-a) in Process 2, and therefore are to be referred tosaid explanation.

The present invention includes, within the scope thereof, cases that theprotected amino group in R¹ and/or the protected carboxy group in R³ aretransformed into free amino group and/or free carboxy group,respectively during the reaction. dioxane, water, etc., or a mixturethereof.

(9) Process 9:

The compound (I-n) or a salt thereof can be prepared by subjecting thecompound (I-m) or a salt thereof to removal reaction of the amino- andcarboxy-protective groups in R_(b) ³.

This reaction is carried out by a conventional method such ashydrolysis, reduction, and the like.

The method of hydrolysis and reduction, and the reaction conditions(e.g. reaction temperature, solvent, etc.) are substantially the same asthose illustrated for removal reaction of the amino-protective group ofthe compound (I-a) in Process 2, and therefore are to be referred tosaid explanation.

In this reaction, the amino- and carboxy-protective groups can beremoved separately or at a time.

(10) Process 10:

The compound (I-p) or a salt thereof can be prepared by subjecting thecompound (I-o) or a salt thereof to removal reaction of the amino- andcarboxy-protective groups in A⁸.

This reaction is carried out by a conventional method such ashydrolysis, reduction, and the like.

The method of hydrolysis and reduction, and the reaction conditions(e.g. reaction temperature, solvent, etc.) are substantially the same asthose illustrated for removal reaction of the amino-protective group ofthe compound (I-a) in Process 2, and therefore are to be referred tosaid explanation.

In this reaction, the amino- and carboxy-protective groups can beremoved separately or at a time.

(11) Process 11:

The compound (I-k) or a salt thereof can be prepared by introducing acarboxy-protective group into the compound (I-l) or a salt thereof.

This reaction is carried out by substantially the same method as thatillustrated for introducing the carboxy-protective group into thecompound (I-f) in Process 5, and therefore, the reaction conditions(e.g. reaction temperature, solvent, etc.) are to be referred to saidexplanation.

(12) Process 12:

The compound (I-r) or a salt thereof can be prepared by reacting thecompound (I-q) or a salt thereof with the compound (VII).

This reaction is usually carried out in a conventional solvent whichdoes not adversely influence the reaction such as tetrahydrofuran,dioxane, water, etc., or a mixture thereof.

The reaction temperature is not critical, and the reaction is usuallycarried out at ambient temperature to under heating.

(13) Process 13:

The compound (I-s) or a salt thereof can be prepared by reacting thecompound (I-r) or a salt thereof with a base.

Suitable base used in this Process may include the same ones as thoseexemplified in Process 1.

This reaction is usually carried out in a conventional solvent whichdoes not adversely influence the reaction such as water, methanol,ethanol, etc., or a mixture thereof.

The reaction temperature is not critical, and the reaction is usuallycarried out under cooling to warming.

The object compounds (I) obtained according to the Processes 1 to 13 asexplained above can be isolated and purified in a conventional manner,for example, extraction, precipitation, fractional crystallization,recrystallization, chromatography, and the like.

Processes A to f for the preparation of the starting compounds (II-d),(II-g), (III-b) to (III-e), (III-g), (III-l), (IV) and (XIV-c) areexplained in detail in the following.

Process A-1:

The compound (II-b) or a salt thereof can be prepared by reacting thecompound (II-a) or a salt thereof with the compound (VIII) or itsreactive derivative at the mercapto group.

Suitable salts of the compounds (II-a) and (II-b) may include the samesalts with a base as exemplified for the compounds (I).

Suitable "reactive derivative at the mercapto group" of the compound(VIII) may include salts with a base as exemplified for the compounds(I).

This reaction is preferably carried out in the presence of a base andsuitable examples thereof are the same as those given in the explanationof Process 1.

The reaction is usually carried out in a conventional solvent which doesnot adversely influence the reaction such as N,N-dimethylformamide,dimethylsulfoxide, methanol, ethanol, chloroform, etc., or a mixturethereof.

The reaction temperature is not critical and the reaction is usuallycarried out at ambient temperature to under warming.

Process A-2:

The compound (II-c) or a salt thereof can be prepared by reducing thecompound (II-b) or a salt thereof.

Suitable salts of the compound (II-c) may include the same salts with abase as exemplified for the compounds (I).

The reducing agent to be used in this reaction may include aconventional one used for conversion of a sulfinyl group to a thio groupsuch as phosphorus halide (e.g. phosphorus trichloride, phosphoruspentachloride, etc.); stannous halide (e.g. stannous chloride, etc.);silicon halide (e.g. silicon tetrachloride, etc.); excess amount of acidhalide such as lower alkanoyl halide (e.g. acetyl bromide, acetylchloride, etc.); combination of alkali metal halide (e.g. sodium iodide,etc.) and acid anhydride such as halo(lower)alkanoic anhydride (e.g.trifluoroacetic anhydride, etc.); and the like.

The reaction is usually carried out in the presence of an acid scavengersuch as lower alkene (e.g. 2-methyl 2-butene, etc.), lower alkyleneoxide (e.g. ethylene oxide, propylene oxide, etc.) and the like.

The reaction is usually carried out in a conventional solvent which doesnot adversely influence the reaction such as chloroform, methylenechloride, tetrahydrofuran, benzene, etc., or a mixture thereof.

The reaction temperature is not critical and the reaction is usuallycarried out under cooling to warming.

Process A-3:

The compound (II-d) or a salt thereof can be prepared by subjecting thecompound (II-c) or a salt thereof to removal reaction of theamino-protective group for R^(a).

Suitable salts of the compound (II-d) may include the same ones asexemplified for the compounds (I).

Suitable method for this removal reaction may include conventional onesuch as a combined method comprising iminohalogenation andiminoetherification, optionally followed by hydrolysis, and the like.

The first and second steps of this method are preferably carried out inan anhydrous solvent. Suitable solvent for the first step (i.e.iminohalogenation) is an aprotic solvent such as methylene chloride,chloroform, diethyl ether, tetrahydrofuran, dioxane, etc., and for thesecond step (i.e. iminoetherification) is usually the same as those inthe above first step. These two steps are usually conducted undercooling. These two steps and the last step (i.e. hydrolysis step) aremost preferably conducted in one-batch system.

Suitable iminohalogenating agents include a halogenating agent such asphosphorus halo compound (e.g. phosphorus trichloride, phosphoruspentachloride, phosphorus tribromide, phosphorus pentabromide,phosphorus oxychloride, etc.), thionyl chloride, phosgene, and the like.

Suitable iminoetherifying agent may be an alcohol such as an alkanol(e.g. methanol, ethanol, propanol, isopropanol, butanol, etc.) or thecorresponding alkanol having alkoxy (e.g. 2-methoxyethanol,2-ethoxyethanol, etc.), and alkoxide of metal such as alkali metal,alkaline earth metal (e.g. sodium methoxide, potassium ethoxide,magnesium ethoxide, lithium methoxide, etc.), and the like.

Thus obtained reaction product is, if necessary, hydrolyzed in aconventional manner.

The hydrolysis is preferably carried out at ambient temperature or undercooling, and proceeds simply pouring the reaction mixture into water ora hydrophilic solvent such as alcohol (e.g. methanol, ethanol, etc.)moistened or admixed with water, and if necessary, with addition of anacid or base as exemplified in Processes 1 and 2.

Process B-1:

The compound (III-b) can be prepared by reacting the compound (III-a)with the compound (IX) or a salt thereof.

Suitable salt of the compound (IX) may include the same one asexemplified for the compounds (I).

This reaction is preferably carried out in the presence of a base asexemplified in Process 1.

This reaction is usually carried out in a conventional solvent whichdoes not adversely influence the reaction such as water, dioxane,tetrahydrofuran, etc., or a mixture thereof.

The reaction temperature is not critical and the reaction is usuallycarried out under cooling to warming.

Process B-2:

The compound (III-c) or a salt thereof can be prepared by reducing thecompound (III-b).

Suitable salts of the compound (III-c) may include the same acidaddition salt as exemplified for the compounds (I).

The reduction can be carried out by a conventional method such aschemical reduction, catalytic reduction, and the like.

The method of chemical reduction and catalytic reduction, and thereaction conditions (e.g. reaction temperature, solvent, etc.) aresubstantially the same as those illustrated for Process 2, and thereforeare to be referred to said explanation.

Process B-3:

The compound (III-d) can be prepared by introducing an amino-protectivegroup into the compound (III-c) or a salt thereof.

The introducing agent of an amino-protective group to be used in thisreaction may include a conventional acylating agent such as thecorresponding acid to the acyl group as aforementioned or its reactivederivative (e.g. acid halide, acid anhydride, etc.), 2-loweralkoxycarbonyloxyimino-2-phenylacetonitrile (e.g.2-tert-butoxycarbonyloxyimino-2-phenylacetonitrile, etc.), alkyl ketonesubstituted by lower alkoxycarbonyl (e.g. lower alkyl acetoacetate, forexample, methyl acetoacetate, etc., etc.), and the like.

This reaction is usually carried out in a conventional solvent whichdoes not adversely influence the reaction such as water, methanol,ethanol, propanol, tetrahydrofuran, dioxane, etc., or a mixture thereof.

This reaction is preferably carried out in the presence of a base, andsuitable examples thereof are the same as those given in the explanationof Process 1.

The reaction temperature is not critical and the reaction is usuallycarried out under cooling to warming.

Process B-4:

The compound (III-e) or a salt thereof can be prepared by subjecting thecompound (III-d) to removal reaction of the carboxy-protective group.

Suitable salts of the compound (III-e) may include the same salts with abase as exemplified for the compounds (I).

This reaction is carried out by a conventional method such ashydrolysis, reduction, and the like.

The method of hydrolysis and reduction, and the reaction conditions(e.g. reaction temperature, solvent, etc.) are substantially the same asthose illustrated for the removal reaction of the amino-protective groupin Process 2, and therefore are to be referred to said explanation.Additionally, hydrolysis can be carried out in the presence of a base,and suitable examples thereof are the same as those in the explanationof Process 1.

Process B-5:

The compound (III-g) or a salt thereof can be prepared by subjecting thecompound (III-f) or a salt thereof to removal reaction of thecarboxy-protective group.

Suitable salts of the compound (III-f) may include the same acidaddition salts as exemplified above, and those of the compound (III-g)may include the same salts as exemplified for the compounds (I).

The reaction is substantially the same as Process B-4, and therefore,the reaction method, reaction conditions (e.g. reaction temperature,solvent, etc.) are to be referred to said explanation.

Process B-6

The compound (III-e) or a salt thereof can be prepared by introducing anamino-protective group into the compound (III-g) or a salt thereof.

This reaction is substantially the same as Process B-3, and therefore,the reaction method and reaction conditions (e.g. reaction temperature,solvent, etc.) are to be referred to said explanation.

Process C:

The compound (IV) or a salt thereof can be prepared by reacting thecompound (IV-a) or its reactive derivative at the amino group or a saltthereof with the compound (X) or its reactive derivative at the carboxygroup or a salt thereof.

Suitable salts of the compound (IV-a) may include the same ones asexemplified for the compounds (I), and those of the compound (X) mayinclude the same salts with a base as exemplified above.

The reaction is substantially the same method as Process 1, andaccordingly, the method, reaction conditions (e.g. reaction temperature,solvent, base, etc.) are to be referred to said explanation.

In this process, carbonyl equivalents, for example, acetal of thecompound (X), wherein A⁵ is lower alkylene having oxo, can also be usedin this reaction and such acetal can easily be transformed into the oxogroup by a conventional method (e.g. hydrolysis, etc.) after thereaction.

Process D-1:

The compound (II-f) or a salt thereof can be prepared by reacting thecompound (II-e) or a reactive derivative at the carboxy group or a saltthereof with lower alkanol substituted by protected amino and protectedcarboxy groups (XII).

Suitable salts of the compounds (II-e) and (II-f) may include the sameones as exemplified for the compounds (I).

Suitable reactive derivative at the carboxy group of the compound (II-e)may include the same ones as the compound (III) in Process 1.

This reaction is carried out by substantially the same method as thatillustrated for introducing the carboxy-protective group into thecompound (I-f) in Process 5, and therefore, the reaction conditions(e.g. reaction temperature, solvent, etc.) are to be referred to saidexplanation.

Process D-2:

The compound (II-g) or a salt thereof can be prepared by subjecting thecompound (II-f) or a salt thereof to removal reaction of theamino-protective group for R^(a).

Suitable salt of the compound (II-g) may include the same ones asexemplified for the compounds (I).

This reaction is carried out by substantially the same method as thatillustrated for removal reaction of the amino-protective group of thecompound (II-c) in Process A-3, and therefore, the reaction conditions(e.g. reaction temperature, solvent, etc.) are to be referred to saidexplanation.

Process E-1:

The compound (III-i) can be prepared by reacting the compound (III-h)with hydroxylamine or a salt thereof.

Suitable salt of hydroxylamine may include the same acid addition saltas exemplified for the compounds (I).

In case that the salt of hydroxylamine is used as the reagent, thereaction can usually be carried out in the presence of a base such asthose illustrated in Process 1.

The reaction is usually carried out in a conventional solvent which doesnot adversely influence the reaction such as methanol, ethanol, etc., ora mixture thereof.

The reaction temperature is not critical and the reaction is usuallycarried out under cooling to warming.

Process E-2:

The compound (III-j) can be prepared by reacting the compound (III-i)with the compound (XIII) or a reactive derivative at the carboxy groupor a salt thereof.

Suitable salt of the compound (XIII) may include the same salt with abase as exemplified for the compounds (I).

Suitable reactive derivative at the carboxy group of the compound (XIII)may include the same ones as illustrated for the compound (III) inProcess 1.

This reaction is carried out by substantially the same method as Process1, and therefore, the reaction conditions (e.g. reaction temperature,solvent, etc.) are to be referred to said explanation.

Process E-3:

The compound (III-k) or a salt thereof can be prepared by reacting thecompound (III-j) or a salt thereof with ammonia.

Suitable salt of the compound (III-k) may include the same acid additionsalt as exemplified for the compounds (I).

This reaction can be carried out in the absence of or in the presence ofa solvent which does not adversely influence the reaction such asdioxane, etc., and the reaction is usually carried out in the absence ofa solvent.

The reaction temperature is not critical and the reaction is usuallycarried out under cooling to warming.

In case that the compound (III-k) is one of the geometrical isomers, itcan be transformed into the other isomer in a conventional manner.

Process E-4:

The compound (III-l) or a salt thereof can be prepared by subjecting thecompound (III-k) or a salt thereof to removal reaction of thecarboxy-protective group for R^(c).

Suitable salt of the compound (III-l) may include the same ones asexemplified for the compounds (I).

This reaction is carried out by a conventional method such ashydrolysis, reduction, and the like.

The method of hydrolysis and reduction, and the reaction conditions(e.g. reaction temperature, solvent, etc.) are substantially the same asthose illustrated for the removal reaction of the amino-protective groupof the compound (I-a) in Process 2, and therefore are to be referred tosaid explanation.

Process F-1:

The compound (XIV-b) can be prepared by reacting the compound (XIV-a) ora reactive derivative at the hydroxy group with N-hydroxyphthalimide.

Suitable reactive derivative at the hydroxy group may include halidesuch as chloride, bromide, and the like.

This reaction is preferably carried out in the presence of a base asexemplified in Process 1.

In case that the compound (XIV-a) is used in a free form, the reactioncan usually be carried out in the presence of a condensing agent asexemplified in Process 1.

This reaction is usually carried out in a conventional solvent whichdoes not adversely influence the reaction such as tetrahydrofuran,N,N-dimethylformamide, etc., or a mixture thereof.

The reaction temperature is not critical and the reaction is usuallycarried out under cooling to warming.

Process F-2:

The compound (XIV-c) or a salt thereof can be prepared by subjecting thecompound (XIV-b) to removal reaction of the phthaloyl group.

Suitable salt of the compound (XIV-c) may include the same acid additionsalt as exemplified for the compounds (I).

This reaction is carried out by a conventional method such ashydrolysis, and the like.

The method of hydrolysis, and the reaction conditions (e.g. reactiontemperature, solvent, etc.) are substantially the same as thoseillustrated for removal reaction of the amino-protective group of thecompound (I-a) in Process 2, and therefore are to be referred to saidexplanation.

The starting compounds (II-d), (II-g), (III-b) to (III-e), (III-l), (IV)and (XIV-c) thus prepared can be isolated in a conventional manner asmentioned for the object compounds of the present invention.

It is to be noted that, in the aforementioned reactions in Processes 1to 13 and A to F or the post-treatment of the reaction mixture therein,in case that the starting or object compounds possess an optical and/orgeometrical isomer(s), it may occasionally be transformed into the otheroptical and/or geometrical isomer(s), and such cases are also includedwithin the scope of the present invention.

In case that the object compounds (I) have a free carboxy group or freeamino group at the 4th or 7th position thereof, it may be transformedinto its pharmaceutically acceptable salts by a conventional method.

The object compounds (I) and the pharmaceutically acceptable saltsthereof of the present invention are novel and exhibit highantimicrobial activity, inhibiting the growth of a wide variety ofpathogenic microorganisms including Gram-positive and Gram-negativemicroorganisms and are useful as antimicrobial agents, especially fororal administration.

Now in order to show the utility of the object compounds (I), the testdata on the in vitro antimicrobial activity of some representativecompounds (I) of this invention are shown in the following.

Test: In vitro Antimicrobial Activity. Test Compounds

No. 1:7-[2-(2-Aminothiazol-4-yl)-DL-glycolamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid (hereinafter referred to as Compound A)

No. 2:7-[2-(3-Methanesulfonamidophenyl)-D-glycinamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid trifluoroacetate (hereinafter referred to as Compound B)

No. 3:7-[2-(3-Methanesulfonamidophenyl)-D-glycinamido]-3-allylthiomethyl-3-cephem-4-carboxylicacid trifluoroacetate (hereinafter referred to as Compound C)

No. 4:7-[2-(3-Methanesulfonamidophenyl)-D-glycinamido]-3-methoxymethyl-3-cephem-4-carboxlicacid (hereinafter referred to as Compound D)

Test Method

In vitro Antimicrobial activity was determined by the two-foldagar-plate dilution method as described below.

One loopful of an overnight culture of each test strain inTripticase-soy broth (approximately 10⁸ viable cells per ml) wasstreaked on heart infusion agar (HIagar) containing gradedconcentrations of antimicrobial agents, and the minimal inhibitoryconcentration (MIC) was expressed in term of μg/ml after incubation at37° C. for 20 hours.

Test Results 1

    ______________________________________                                        MIC (μg/ml)                                                                           Microorganisms                                                                  Staphylococcus                                                                             Batilus subtilis                                    Test compounds                                                                             aureus 209P JC-1                                                                           ATCC 6633                                           ______________________________________                                        A            1.56         0.78                                                B            1.56         0.10                                                C            3.13         0.78                                                D            1.56         0.39                                                ______________________________________                                    

For therapeutic administration, the object compounds (I) and thepharmaceutically acceptable salts thereof of the present invention areused in the form of conventional pharmaceutical preparation whichcontains said compound, as active ingredients, in admixture withpharmaceutically acceptable carriers such as an organic or inorganicsolid or liquid excipient which is suitable for oral, parenteral andexternal administration. The pharmaceutical preparations may be in solidform such as tablet, granule, powder, capsule, or liquid form such assolution, suspension, syrup, emulsion, lemonade and the like.

If needed, there may be included in the above preparations auxiliarysubstances, stabilizing agents, wetting agents and other commonly usedadditives such as lactose, magnesium stearate, terra alba, sucrose, cornstarch, talc, stearic acid, gelatin, agar, pectin, peanut oil, oliveoil, cacao butter, ethyleneglycol and the like.

While the dosage of the compounds (I) may vary from and also depend uponthe age, conditions of the patient, a kind of diseases, a kind of thecompounds (I) to be applied, etc. In general, amounts between 1 mg andabout 4,000 mg or even more per day may be administered to a patient. Anaverage single dose of about 50 mg, 100 mg, 250 mg, 500 mg, 1000 mg,2000 mg of the object compounds (I) of the present invention may be usedin treating diseases infected by pathogenic microorganisms.

The following examples are given for the purpose of illustrating thepresent invention.

PREPARATION OF THE STARTING COMPOUNDS Preparation 1

To a solution of benzhydryl7-(2-phenylacetamido)-3-chloromethyl-3-cephem-4-carboxylate-1-oxide (25g) in N,N-dimethylformamide (150 ml) were added triethylamine (6.42 ml)and 2-propene-1-thiol (8.0 ml), and the mixture was stirred at 25° C.for 3 hours. The reaction mixture was poured into a saturated aqueoussolution of sodium chloride (1.5 l), followed by collecting theprecipitated solid by filtration, which was washed with water anddiisopropyl ether, and then dried to give benzhydryl7-(2-phenylacetamido)-3-allylthiomethyl-3-cephem-4-carboxylate-1-oxide(24.2 g).

I.R. (Nujol): 1775, 1715, 1644, 1170, 1030 cm⁻¹

NMR δppm (DMSO-d₆): 3.00 (2H, d, J=7Hz), 3.6 (6H, m), 5.0 (1H, d,J=5Hz), 5.90 (1H, dd, J=5Hz, 8Hz), 4.8-5.6 (3H, m), 7.00 (1H, s), 7.4(15H, s), 8.40 (1H, d, J=8Hz)

Preparation 2

Benzhydryl7-(2-phenylacetamido)-3-methylthiomethyl-3-cephem-4-carboxylate-1-oxide(13.7 g) was obtained by reacting benzhydryl7-(2-phenylacetamido)-3-chloromethyl-3-cephem-4-carboxylate-1-oxide (15g) with 30% methanolic methanethiol (15 ml) in substantially the samemanner as that of Preparation 1.

I.R. (Nujol): 3300, 1775, 1710, 1650, 1172, 1027 cm⁻¹

NMR δppm (DMSO-d₆): 1.80 (3H, s), 3.3-4.0 (6H, m), 5.02 (1H, d, J=5Hz),5.93 (1H, dd, J=5Hz, 8Hz), 7.02 (1H, s), 7.50 (15H, s), 8.40 (1H, d,J=8Hz)

Preparation 3

Benzhydryl7-(2-phenylacetamido)-3-ethylthiomethyl-3-cephem-4-carboxylate-1-oxide(14.1 g) was obtained by reacting benzhydryl7-(2-phenylacetamido)-3-chloromethyl-3-cephem-4-carboxylate-1-oxide (15g) with ethanethiol (4.05 ml) in substantially the same manner as thatof Preparation 1.

I.R. (Nujol): 3280, 1776, 1708, 1647, 1172, 1015 cm⁻¹

NMR δppm (DMSO-d₆): 0.95 (3H, t, J=7Hz), 2.28 (2H, q, J=7Hz), 3.5-4.0(6H, m), 5.02 (1H, d, J=5Hz), 5.93 (1H, dd, J=5Hz, 8Hz), 7.02 (1H, s),7.5 (15H, s), 8.43 (1H, d, J=8Hz)

Preparation 4

To a solution of benzhydryl7-(2-phenylacetamido)-3-allylthiomethyl-3-cephem-4-carboxylate-1-oxide(26 g) in methylene chloride (500 ml) was added dropwise phosphorustrichloride (20 ml) at 5° C. with stirring, and the stirring wascontinued for an hour. The mixture was poured into a mixture ofmethylene chloride (200 ml) and water (400 ml), followed by separatingout the organic layer, which was washed twice with an aqueous solutionof sodium chloride (200 ml) and dried over anhydrous magnesium sulfate.After removal of the solvent, the residue was pulverized withdiisopropyl ether to give benzhydryl7-(2-phenylacetamido)-3-allylthiomethyl-3-cephem-4-carboxylate (22 g).

I.R. (Nujol): 1770, 1715, 1650 cm⁻¹

NMR (DMSO-d₆): 3.0 (2H, d, J=7Hz), 3.6 (6H, m), 5.00 (1H, d, J=5Hz),5.67 (1H, dd, J=5Hz, 8Hz), 4.8-5.5 (3H, m), 6.90 (1H, s), 7.40 (15H, m),9.10 (1H, d, J=8Hz)

Preparation 5

To a solution of benzhydryl7-(2-phenylacetamido)-3-methylthiomethyl-3-cephem-4-carboxylate-1-oxide(15.1 g) in methylene chloride (150 ml) was added 2-methyl-2-butene (5.7ml), followed by adding dropwise acetyl bromide (5.2 ml) underice-cooling and stirring for half an hour. After addition of water, themixture was adjusted to pH about 5 with an aqueous solution of sodiumbicarbonate, washed three times with an aqueous solution of sodiumchloride, dried over anhydrous magnesium sulfate and then evaporatedunder reduced pressure to give benzhydryl7-[2-phenylacetamido)-3-methylthiomethyl-3-cephem-4-carboxylate (13.5g).

I.R. (Nujol): 3380, 1785, 1715, 1652 cm⁻¹

NMR δppm (DMSO-d₆): 1.83 (3H, s), 3.60 (4H, broad s), 3.66 (2H, broads), 5.23 (1H, d, J=5Hz), 5.75 (1H, dd, J=5Hz, 8Hz), 6.97 (1H, s), 7.43(15H, s), 9.17 (1H, d, J=8Hz)

Preparation 6

Benzhydryl7-(2-phenylacetamido)-3-ethylthiomethyl-3-cephem-4-carboxylate (23 g)was obtained by reacting benzhydryl7-(2-phenylacetamido)-3-ethylthiomethyl-3-cephem-4-carboxylate-1-oxide(30 g) with acetyl bromide (10.2 ml) in the presence of2-methyl-2-butene (11.1 ml) in substantially the same manner as that ofPreparation 5.

I.R. (Nujol): 3300, 1772, 1701, 1650 cm⁻¹

NMR δppm (DMSO-d₆): 1.00 (3H, t, J=7Hz), 2.33 (2H, q, J=7Hz), 3.56 (6H,broad s), 5.17 (1H, d, J=5Hz), 5.76 (1H, dd, J=5Hz, 8Hz), 7.00 (1H, s),9.13 (1H, d, J=8Hz)

Preparation 7

To a suspension of phosphorus pentachloride (16.1 g) and pyridine (6.3ml) in methylene chloride (100 ml) were added benzhydryl7-(2-phenylacetamido)-3-allylthiomethyl-3-cephem-4-carboxylate (22 g)and methylene chloride (100 ml) at 5° C., and the mixture was stirred atthe same temperature for an hour. After cooling to -20° C., methanol (10ml) was added thereto, followed by stirring at -10° C. for half an hour.To this mixture was added water (10 ml) and stirred for 10 minutes. Tothe separated methylene chloride was added an aqueous solution of sodiumbicarbonate until the pH value of the aqueous solution became 5.0, andthe mixture was shaken. After separating out the organic layer, it waswashed with an aqueous solution of sodium chloride, dried over anhydrousmagnesium sulfate and evaporated to dryness. The residue obtained waspulverized with diisopropyl ether to give benzhydryl7-amino-3-allylthiomethyl-3-cephem-4-carboxylate (6.5 g).

I.R. (Nujol): 1770, 1710 cm⁻¹

NMR δppm (DMSO-d₆): 2.93 (2H, d, J=7Hz), 3.3-3.7 (4H, m), 5.00 (1H, d,J=5Hz), 5.60 (1H, d, J=5Hz), 4.5-5.6 (3H, m), 6.91 (1H, s), 7.5 (10H, m)

Preparation 8

Benzhydryl 7-amino-3-methylthiomethyl-3-cephem-4-carboxylate (5.0 g) wasobtained by reacting benzhydryl7-(2-phenylacetamido)-3-methylthiomethyl-3-cephem-4-carboxylate (13.5 g)with phosphorus pentachloride (7.74 g), pyridine (3 ml) and methanol(100 ml) in substantially the same manner as that of Preparation 7.

I.R. (Nujol): 1765, 1725 cm⁻¹

NMR δppm (DMSO-d₆): 1.81 (3H, s), 3.52 (2H, broad s), 3.60 (2H, broads), 4.83,5.13 (2H, ABq, J=5Hz), 6.97 (1H, s), 7.40 (10H, s)

Preparation 9

Benzhydryl 7-amino-3-ethylthiomethyl-3-cephem-4-carboxylate (10.0 g) wasobtained by reacting benzhydryl7-(2-phenylacetamido)-3-ethylthiomethyl-3-cephem-4-carboxylate (23.0 g)with phosphorus pentachloride (12.90 g), pyridine (5.0 ml) and methanol(165 ml) in substantially the same manner as that of Preparation 7.

I.R. (Nujol): 1770, 1720 cm⁻¹

NMR δppm (DMSO-d₆): 0.96 (3H, t, J=7Hz), 2.30 (2H, q, J=7Hz), 3.50 (2H,broad s), 3.60 (2H, broad s), 4.80,5.17 (2H, ABq, J=5Hz), 7.00 (1H, s),7.43 (10H, s)

Preparation 10

To a solution of diketene (1.3 ml) in methylene chloride (10 ml) wasadded dropwise a solution of bromine (1.24 g) in methylene chloride (10ml) at -30° C. with stirring, and the stirring was continued at -20° C.for half an hour to prepare a solution of 4-bromoacetoacetyl bromide.This solution was added dropwise to a solution of benzhydryl7-amino-3-methylthiomethyl-3-cephem-4-carboxylate (4.44 g) andtrimethylsilylacetamide (5.46 g) in methylene chloride (100 ml) at -30°to -20° C. over a period of 5 minutes with stirring, and the stirringwas continued at -10° C. for half an hour. After addition of water, theresultant mixture was adjusted to pH 7.5 with an aqueous solution ofsodium bicarbonate, followed by separating out the organic layer, whichwas washed with water and an aqueous solution of sodium chloride, driedover anhydrous magnesium sulfate, and then evaporated to dryness to givebenzhydryl 7-(4-bromoacetoacetamido)-3-methylthiomethyl-3-cephem-4-carboxylate (6.0g).

IR (Nujol): 1770, 1710, 1625 cm⁻¹

NMR δppm (DMSO-d₆): 1.77 (3H, s), 3.6 (6H, m), 4.33 (2H, s), 5.15 (1H,d, J=4Hz), 5.73 (1H, dd, J=4Hz, 8Hz), 6.86 (1H, s), 7.3 (10H, m), 9.1(1H, d, J=8Hz)

Preparation 11

To a solution of ethyl 2-(2-formamidothiazol-4-yl)-2-methoxyiminoaceticacid (syn isomer) (19 g) in methanol (200 ml) were added 50% formic acid(200 ml) and zinc (29 g), and the mixture was stirred at 5° to 10° C.for 6 hours. After filtration, the reaction mixture was evaporated,followed by dissolving the residue in water (150 ml). The resultantaqueous solution was adjusted to pH 6.5 with 4N aqueous solution ofsodium hydroxide, followed by addition of ethanol (150 ml),2-tert-butoxycarbonyloxyimino-2-phenylacetonitrile (18.2 g) andtriethylamine (8.0 g). After stirring at ambient temperature for 24hours, the reaction mixture was filtered, followed by removal of theorganic solvent. The remained aqueous solution was washed with ethylacetate, adjusted to pH 4 with 10% hydrochloric acid and then extractedwith ethyl acetate. The extract was washed with an aqueous solution ofsodium chloride, dried over anhydrous magnesium sulfate and evaporatedto dryness under reduced pressure to give a residue, which was washedwith diethyl ether to obtainN-tert-butoxycarbonyl-2-(2-formamidothiazol-4-yl)glycine (3.3 g).

IR (Nujol): 3250, 3180, 1720, 1700, 1670, 1640, 1540, 1510 cm⁻¹

NMR δppm (DMSO-d₆): 1.40 (9H, s), 5.18 (1H, d, J=8Hz), 7.17 (1H, s),8.43 (1H, s)

Preparation 12

Bromoacetic acid (10.45 g) was dissolved in methanol (30 ml). To thesolution was added an equivalent volume of diphenyl diazomethane inethyl acetate at 45° C., and the reaction mixture was stirred at thesame temperature for an hour. The solution was washed with 5% aqueoussodium bicarbonate and a saturated aqueous sodium chloride, and thendried over magnesium sulfate. The solution was evaporated in vacuo togive an oily product. This oil was dissolved in N,N-dimethylformamide(60 ml). To the solution was added N-hydroxyphthalimide (11.7 g) andtriethylamine (15.1 ml), and the reaction mixture was stirred at ambienttemperature for an hour. The resultant mixture was poured into asaturated aqueous sodium chloride (500 ml). The precipitates werecollected by filtration, washed with water, and then dissolved inmethylene chloride (700 ml). The solution was washed with a saturatedaqueous sodium chloride, dried over magnesium sulfate, and thenevaporated under reduced pressure to give benzhydryl2-phthalimidooxyacetate (20.4 g), mp 173°-175° C.

IR (Nujol): 1754, 1730 cm⁻¹

NMR δppm (CDCl₃,δ): 4.93 (2H, s), 7.0 (1H, s), 7.3 (10H, s), 7.73 (4H,s).

Preparation 13

To a solution of benzhydryl 2-phthalimidooxyacetate (10 g) in methylenechloride (100 ml) was added a solution of hydrazine hydrate (6.08 g) inmethanol (7 ml). The reaction mixture was stirred at ambient temperaturefor an hour. The precipitates were collected by filtration and washedwith methylene chloride. The filtrate and the washings were combined,adjusted to pH 7.0 with conc. hydrochloric acid, and washed with asaturated aqueous sodium chloride, and then dried over magnesiumsulfate. The solution was evaporated in vacuo to give benzhydryl2-aminooxyacetate (6.0 g).

IR (film): 3320, 1750 cm⁻¹

NMR δppm (CDCl₃,δ): 4.33 (2H, s), 5.86 (2H, broad s), 7.00 (1H, s), 7.3(10H, s)

Preparation 14

To a suspension of (2-formamidothiazol-4-yl)-glyoxylic acid (6.0 g) inwater (60 ml) and pyridine (6 ml) was added a solution of benzhydryl2-aminooxyacetate (9.0 g) in tetrahydrofuran (40 ml). The reactionmixture was stirred at ambient temperature for 3 hours. To the resultantsolution was added ethyl acetate (200 ml). The separated organic layerwas washed with 5% hydrochloric acid (100 ml), a saturated aqueoussodium bicarbonate and a saturated aqueous sodium chloride, and thendried over magnesium sulfate. The solvent was distilled off to give2-(2-formamidothiazol-4-yl)-2-benzhydryloxycarbonylmethoxyiminoaceticacid (syn isomer) (13.0 g), mp 143°-151° C.

IR (Nujol): 3150, 1733, 1692 cm⁻¹

NMR δppm (DMSO-d₆): 5.0 (2H, broad s), 6.97 (1H, s), 7.40 (20H, m), 7.56(1H, s), 8.60 (1H, s), 12.77 (1H, broad s)

Preparation 15

To a solution of2-benzhydryloxycarbonylmethoxyimino-2-(2-formamidothiazol-4-yl)aceticacid (syn isomer) (7.5 g) in tetrahydrofuran (40 ml) was addedtrifluoroacetic anhydride (7.9 g) at -16° C. for 10 minutes. To thereaction mixture was added triethylamine (5.3 ml) at -10° C., and thenthe mixture was stirred for 90 min at 0° to 5° C. To the above mixturewere added ethyl acetate (100 ml) and water (100 ml), and adjusted to pH7.5 with a saturated aqueous sodium bicarbonate. The aqueous layer wasadjusted to pH 2.0 with conc. hydrochloric acid and extracted with ethylacetate (200 ml). The organic layer was washed with a saturated aqueoussodium chloride and dried over magnesium sulfate. The solvent wasremoved by evaporation under reduced pressure to give an oil. n-Hexanewas added to the oil and the precipitated substance was collected byfiltration to give crystalline2-benzhydryloxycarbonylmethoxyimino-2-[2-(2,2,2-trifluoroacetamido)thiazol-4-yl]aceticacid (syn isomer) (6.0 g), mp 178°-180° C.

IR (Nujol): 1752, 1726 cm⁻¹

NMR δppm (DMSO-d₆): 4.98 (2H, s), 6.92 (1H, s), 7.32 (10H, m), 7.69 (1H,s)

PREPARATION OF THE OBJECT COMPOUNDS EXAMPLE 1

To a solution ofN-tert-butoxycarbonyl-2-(3-methanesulfonamidophenyl)-D-glycine (3.3 g)and triethylamine (1.34 ml) in tetrahydrofuran (50 ml) was addeddropwise a solution of ethyl chloroformate (0.91 ml) in tetrahydrofuran(10 ml) at -10° to -7° C. with stirring, and the stirring was continuedat the same temperature for 40 minutes to give a solution of theactivated acid. This solution was added dropwise to a solution ofbenzhydryl 7-amino-3-methylthiomethyl-3-cephem-4-carboxylate (3.0 g) inmethylene chloride (100 ml) at -30° C. over a period of 5 minutes withstirring, and the stirring was continued at the same temperature for anhour. After addition of water, the reaction mixture was stirred for halfan hour, followed by extracting with methylene chloride (100 ml). Theextract was washed twice with 5% aqueous solution of sodium bicarbonate(50 ml) and an aqueous solution of sodium chloride, and then dried overanhydrous magnesium sulfate, followed by evaporation under reducedpressure to give benzhydryl7-[N-tert-butoxycarbonyl-2-(3-methanesulfonamidophenyl)-D-glycinamido]-3-methylthiomethyl-3-cephem-4-carboxylate(6.5 g).

I.R. (Nujol): 1780, 1710, 1680, 1152 cm⁻¹

NMR δppm (DMSO-d₆): 1.36 (9H, s), 1.76 (3H, s), 2.98 (3H, s), 3.3-3.8(4H, m), 5.12 (1H, d, J=5Hz), 5.73 (1H, dd, J=5Hz, 8Hz), 6.93 (1H, s),7.1-7.4 (14H, m), 9.93 (1H, d, J=8Hz)

EXAMPLE 2

Benzhydryl7-[N-tert-butoxycarbonyl-2-(3-methanesulfonamidophenyl)-D-glycinamido]-3-ethylthiomethyl-3-cephem-4-carboxylate(5.4 g) was obtained by reacting benzhydryl7-amino-3-ethylthiomethyl-3-cephem-4-carboxylate (3.08 g) withN-tert-butoxycarbonyl-2-(3-methanesulfonamidophenyl)-D-glycine (3.3 g)in substantially the same manner as that of Example 1.

I.R. (Nujol): 3250, 1780, 1700, 1530, 1490, 1455 cm⁻¹

NMR δppm (DMSO-d₆): 0.98 (3H, t, J=7Hz), 1.38 (9H, s), 2.28 (2H, q,J=7Hz), 2.97 (3H, s), 3.4-3.8 (4H, m), 5.12 (1H, d, J=5Hz), 5.24 (1H,m), 5.73 (1H, dd, J=5Hz, 7Hz), 6.93 (1H, s), 7.0-7.7 (14H, m), 9.89 (1H,d, J=8Hz), 10.43 (1H, s)

EXAMPLE 3

(1) To a suspension of tert-butyl7-amino-3-methoxymethyl-3-cephem-4-carboxylate tosylate (13.1 g) inmethylene chloride (200 ml) was added water (100 ml), followed byadjusting to pH about 6 with an aqueous solution of sodium bicarbonate.After separating out the methylene chloride layer, the remaining aqueoussolution was extracted with methylene chloride (50 ml). The combinedmethylene chloride solution was washed with an aqueous solution ofsodium chloride, dried over anhydrous magnesium sulfate and then treatedwith an activated charcoal, followed by evaporation under reducedpressure to give tert-butyl7-amino-3-methoxymethyl-3-cephem-4-carboxylate (5.7 g).

I.R. (Nujol): 3370, 1760, 1740, 1710 cm⁻¹

NMR δppm (DMSO-d₆): 1.5 (9H, s), 2.3 (2H, broad s), 3.24 (3H, s), 3.51(2H, s), 4.15 (2H, s), 4.8 (1H, d, J=5Hz), 5.04 (1H, d, J=5Hz)

(2) On the other hand, to a solution ofN-tert-butoxycarbonyl-2-(3-methanesulfonamidophenyl)-D-glycine (8.89 g)and triethylamine (2.61 g) in dry tetrahydrofuran (95 ml) was addeddropwise a solution of ethyl chloroformate (2.8 g) in drytetrahydrofuran (25 ml) at -10° to -7° C. over a period of 10 minuteswith stirring, and stirring was continued at the same temperature for 40minutes to prepare a solution of the activated acid.

(3) To a solution of the compound (5.65 g) obtained according to Example3-(1) in dry methylene chloride (190 ml) was added dropwise the solutionof the activated acid prepared above at -30° C. over a period of 10minutes with stirring, and the stirring was continued at the sametemperature for an hour. After addition of water (100 ml), the reactionmixture was stirred for half an hour. The methylene chloride layer wasseparated out therefrom and the remaining aqueous solution was extractedwith methylene chloride. The combined methylene chloride solution waswashed twice with 5% aqueous solution of sodium bicarbonate (100 ml) andan aqueous solution of sodium chloride, dried over anhydrous magnesiumsulfate, and then treated with an activated charcoal, followed byevaporation under reduced pressure to give tert-butyl7-[N-tert-butoxycarbonyl-2-(3-methanesulfonamidophenyl)-D-glycinamido]-3-methoxymethyl-3-cephem-4-carboxylate(17.8 g).

I.R. (Nujol): 3250, 1780, 1710, 1680 cm⁻¹

NMR δppm (DMSO-d₆): 1.36 (9H, s), 1.46 (9H, s), 2.97 (3H, s), 3.18 (3H,s), 3.4 (2H, broad s), 4.07 (2H, broad s), 5.03 (1H, d, J=5Hz), 5.24(1H, d, J=8Hz), 5.73 (1H, dd, J=5Hz, 8Hz), 6.93-7.43 (4H, m), 7.5 (1H,d, J=8Hz), 9.16 (1H, d, J=8Hz), 9.73 (1H, broad s),

The compounds described in the following Examples 4 to 7 were obtainedby reacting the corresponding 7-aminocephalosporanic acid derivativewith 2-(3-methanesulfonamidophenyl)-D-glycine in substantially the samemanner as that of Example 3.

EXAMPLE 4

7-[2-(3-Methanesulfonamidophenyl)-D-glycinamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid.

I.R. (Nujol): 1758, 1687 (shoulder), 1666, 1144, 974 cm⁻¹

EXAMPLE 5

7-[2-(3-Methanesulfonamidophenyl)-D-glycinamido]-3-ethylthiomethyl-3-cephem-4-carboxylicacid.

I.R. (Nujol): 3500, 3150, 1780, 1685, 1460 cm⁻¹

EXAMPLE 6

7-[2-(3-Methanesulfonamidophenyl)-D-glycinamido]-3-methoxymethyl-3-cephem-4-carboxylicacid.

I.R. (Nujol): 3500, 3150, 1760, 1685 cm⁻¹

EXAMPLE 7

7-[2-(3-Methanesulfonamidophenyl)-D-glycinamido]-3-allylthiomethyl-3-cephem-4-carboxylicacid trifluoroacetate.

I.R. (Nujol): 1760, 1680, 1600, 1140 cm⁻¹

EXAMPLE 8

A mixture of benzhydryl7-[N-tert-butoxycarbonyl-2-(3-methanesulfonamidophenyl)-D-glycinamido]-3-methylthiomethyl-3-cephem-4-carboxylate(6.5 g), anisole (5.0 ml) in trifluoroacetic acid (20 ml) was stirred at25° C. for 15 minutes. After the reaction, the reaction mixture wasadded dropwise to diisopropyl ether (300 ml), and the precipitated solidwas collected by filtration, washed with diisopropyl ether and thendissolved in a mixture of water (50 ml) and ethyl acetate (50 ml). Afterthe aqueous layer was separated out, it was washed with ethyl acetate,followed by removal of ethyl acetate from the aqueous solutioncompletely under reduced pressure. The resultant aqueous solution wasadjusted to pH 3.8 with an aqueous solution of sodium bicarbonate andsubjected to column chromatography using "Diaion HP-20" (90 ml). Afterwashing with water (180 ml), elution was carried out with 30% isopropylalcohol, and the fractions containing the desired compound werecollected and lyophilized to give7-[2-(3-methanesulfonamidophenyl)-D-glycinamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid (4.8 g).

I.R. (Nujol): 1758, 1687 (shoulder), 1666, 1144, 974 cm⁻¹

NMR δppm (D₂ O+DCl): 1.98 (3H, s), 3.15 (3H, s), 3.45 (2H, broad s),3.56 (2H, broad s), 5.12 (1H, d, J=5Hz), 5.30 (1H, s), 5.70 (1H, d,J=5Hz), 7.45 (4H, s)

EXAMPLE 9

7-[2-(3-Methanesulfonamidophenyl)-D-glycinamido]-3-ethylthiomethyl-3-cephem-4-carboxylicacid (1.6 g) was obtained by reacting benzhydryl7-[N-tert-butoxycarbonyl-2-(3-methanesulfonamidophenyl)-D-glycinamido]-3-ethylthiomethyl-3-cephem-4-carboxylate(3.7 g) with trifluoroacetic acid (7.4 ml) in the presence of anisole(7.4 ml) in substantially the same manner as that of Example 8, mp 188°C. (dec.).

I.R. (Nujol): 3500, 3150, 1780, 1685, 1460 cm⁻¹

NMR δppm (D₂ O+DCl): 1.13 (3H, t, J=7Hz), 2.48 (2H, q, J=7Hz), 3.11 (3H,s), 3.3-3.8 (4H, m), 5.30 (1H, s), 5.65 (1H, d, J=5Hz), 7.43 (4H, s)

EXAMPLE 10

To a solution of tert-butyl7-[N-tert-butoxycarbonyl-2-(3-methanesulfonamidophenyl)-D-glycinamido]-3-methoxymethyl-3-cephem-4-carboxylate(15 g) in anisole (11.25 ml) was added dropwise trifluoroacetic acid(33.75 ml) below 15° C. over a period of 10 minutes with stirring, andthe stirring was continued at 15° to 20° C. for half an hour. Thereaction mixture was poured into diisopropyl ether (750 ml), followed bystirring at ambient temperature for 20 minutes. After the precipitatedsolid was collected by filtration and washed with diisopropyl ether, itwas poured into a mixture of ethyl acetate (100 ml) and water (100 ml)and stirred for a while. The aqueous solution was separated outtherefrom, and the remaining organic layer was extracted with water. Thecombined aqueous solution was concentrated under reduced pressure, andthe concentrate was adjusted to pH about 3.8 with an aqueous solution ofsodium bicarbonate, followed by subjecting to column chromatographyusing non-ionic absorption resin "Diaion HP-20" (225 ml), which waswashed with water (450 ml) and then eluted with 30% isopropyl alcohol.The fractions containing the desired compound were collected andevaporated under reduced pressure, and the residue was lyophilized togive7-[2-(3-methanesulfonamidophenyl)-D-glycinamido]-3-methoxymethyl-3-cephem-4-carboxylicacid (4.45 g).

I.R. (Nujol): 3500, 3150, 1760, 1685 cm⁻¹

NMR δppm (D₂ O+DCl): 3.13 (3H, s), 3.26 (3H, s), 3.42 (2H, q, J=18Hz),4.25 (2H, s), 5.06 (1H, d, J=5Hz), 5.27 (1H, s), 5.73 (1H, d, J=5Hz),7.42 (4H, s)

EXAMPLE 11

Benzhydryl 7-amino-3-allylthiomethyl-3-cephem-4-carboxylate (3.3 g) andN-tert-butoxycarbonyl-2-(3-methanesulfonamidophenyl)-D-glycine (3.44 g)were treated in substantially the same manner at that of Example 1 togive an oily product (6.5 g). A mixture of this oil, trifluoroaceticacid (15 ml) and anisole (15 ml) was stirred at 20° C. for half an hour,followed by adding dropwise to diisopropyl ether. The presipitated solidwas collected by filtration, washed with diisopropyl ether and thendissolved in a mixture of water (50 ml) and ethyl acetate (50 ml). Theaqueous layer was separated out and washed with diethyl ether, followedby removal of the organic solvent therefrom completely. The resultantaqueous solution was lyophilized to give7-[2-(3-methanesulfonamidophenyl)-D-glycinamido]-3-allylthiomethyl-3-cephem-4-carboxylicacid trifluoroacetate (2 g).

I.R. (Nujol): 1760, 1680, 1600, 1140 cm⁻¹

NMR δppm (D₂ O+DCl): 3.20 (2H, m), 3.23 (3H, s), 3.6 (7H, m), 5.2 (1H,d, J=5Hz), 5.45 (1H, s), 5.80 (1H, d, J=5Hz), 7.55 (3H, m)

EXAMPLE 12

To a solution of benzhydryl7-[4-bromoacetoacetamido)-3-methylthiomethyl-3-cephem-4-carboxylate (6.0g) in tetrahydrofuran (30 ml) was added dropwise a solution of thiourea(0.85 g) and sodium bicarbonate (0.94 g) in tetrahydrofuran (30 ml) andwater (24 ml) at 25° C. with stirring, and the stirring was continued at28° to 30° C. for an hour. The reaction mixture was poured into amixture of ethyl acetate (100 ml) and water (100 ml), followed byseparating out the organic layer, which was washed twice with an aqueoussolution of sodium chloride, dried over anhydrous magnesium sulfate andthen evaporated to give benzhydryl7-[2-(2-aminothiazol-4-yl)-acetamido]-3-methylthiomethyl-3-cephem-4-carboxylate(4.0 g).

IR (Nujol): 1773, 1715, 1653 cm⁻¹

NMR δppm (DMSO-d₆): 1.85 (3H, s), 3.3-3.8 (6H, m), 5.25 (1H, d, J=5Hz),5.80 (1H, dd, J=5Hz, 8Hz), 6.32 (1H, s), 7.00 (1H, s), 7.43 (10H, s),8.95 (1H, d, J=8Hz)

EXAMPLE 13

tert-Butyl 7-amino-3-methoxymethyl-3-cephem-4-carboxylate tosylate (2.0g) was dissolved in a mixture of acetone (40 ml) and a saturated aqueoussolution of sodium bicarbonate (15 ml), and thereto was added dropwise asolution of the activated acid, which was prepared from2-(2-formamidothiazol-4-yl)-2-methoxyiminoacetic acid (syn isomer) (1.06g), phosphorus oxychloride (0.85 g) and N,N-dimethylformamide (0.41 g)in tetrahydrofuran (10 ml), at 0° to 5° C. over a period of 10 minutes.During the addition, the pH value of the reaction mixture was maintainedat pH 7.0 to 7.5 with a saturated aqueous solution of sodium carbonate.After stirring for an hour, the reaction mixture was diluted with water(50 ml), followed by extracting twice with ethyl acetate. The combinedextract was washed with 5% aqueous solution of sodium bicarbonate andwater, dried and then evaporated to dryness under reduced pressure togive a residue, which was triturated with diethyl ether to obtaintert-butyl7-[2-(2-formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-methoxymethyl-3-cephem-4-carboxylate(syn isomer) (1.14 g).

IR (Nujol): 3250, 3100, 1790, 1710, 1660 cm⁻¹

NMR δppm (DMSO-d₆): 1.49 (9H, s), 3.21 (3H, s), 3.28 (2H, broad s), 3.89(3H, s), 4.1 (2H, s), 5.16 (1H, d, J=5Hz), 5.80 (1H, dd, J=5Hz, 8Hz),7.36 (1H, s), 8.48 (1H, s), 9.6 (1H, d, J=8Hz), 12.66 (1H, broad s)

EXAMPLE 14

tert-Butyl7-[2-(2-formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-methylthiomethyl-3-cephem-4-carboxylate(syn isomer) (0.91 g) was obtained by reacting tert-butyl7-amino-3-methylthiomethyl-3-cephem-4-carboxylate (0.90 g) with2-(2-formamidothiazol-4-yl)-2-methoxyiminoacetic acid (syn isomer) (0.85g) according to the similar manner to that of Example 13.

IR (Nujol): 3250, 3050, 1780, 1690 cm⁻¹

NMR δppm (DMSO-d₆): 1.47 (9H, s), 1.97 (3H, s), 3.29 (2H, broad s), 3.55(2H, ABq, J=13Hz), 3.87 (3H, s), 5.21 (1H, d, J=5Hz), 5.76 (1H, dd,J=5Hz, 8Hz), 7.38 (1H, s), 8.49 (1H, s), 9.66 (1H, d, J=8Hz), 12.56 (1H,broad s)

The compounds described in the following Examples 15 to 17 were obtainedby reacting the 7-aminocephalosporanic acid derivatives with thecorresponding acid according to the similar manner to that of Example13.

EXAMPLE 15

7-[2-(2-Aminothiazol-4-yl)acetamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid

IR (Nujol): 1763, 1654 cm⁻¹

EXAMPLE 16

7-[2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-methoxymethyl-3-cephem-4-carboxylicacid hydrochloride (syn isomer)

IR (Nujol): 3300, 1780, 1720, 1660, 1640 cm⁻¹

EXAMPLE 17

7-[2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid (syn isomer)

IR (Nujol): 3350, 1770, 1670 cm⁻¹

EXAMPLE 18

The mixture of benzhydryl7-[2-(2-aminothiazol-4-yl)acetamido]-3-methylthiomethyl-3-cephem-4-carboxylate(3.3 g), trifluoroacetic acid (10 ml) and anisole (10 ml) in methylenechloride (10 ml) was stirred at 10° C. for an hour. After benzene (50ml) was added to the reaction mixture, the trifluoroacetic acid thereinwas azeotropically removed under reduced pressure. The remained aqueoussolution was poured into a mixture of ethyl acetate (100 ml) and water(100 ml), followed by adjusting to pH 7.5 with sodium bicarbonate. Afterseparating out the aqueous solution, the organic solvent was removedtherefrom by evaporation completely under reduced pressure, followed byadjusting to pH 3.0 with 10% hydrochloric acid. The precipitated solidwas collected by filtration and then dried to give7-[2-(2-aminothiazol-4-yl)acetamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid (0.95 g).

IR (Nujol): 1763, 1654 cm⁻¹

NMR δppm (DMSO-d₆): 2.01 (3H, s), 3.48 (2H, s), 3.65 (4H, broad s), 5.17(1H, d, J=5Hz), 5.67 (1H, dd, J=5Hz, 8Hz), 6.38 (1H, s), 9.00 (1H, d,J=8Hz)

EXAMPLE 19

To a cold suspension of tert-butyl7-[2-(2-formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-methoxymethyl-3-cephem-4-carboxylate(synisomer)(1.0 g) in methylene chloride (20 ml) were added trifluoroaceticacid (4.4 g) and anisole (0.2 ml), and the mixture was gradually warmedat 40° C., followed by stirring at 10° C. for 3 hours. After evaporationof the reaction mixture, to the residue was added ethyl acetate (20 ml),followed by extracting with an aqueous solution of sodium bicarbonate.The resultant aqueous solution was adjusted to pH 2.0 with dilutedhydrochloric acid and then extracted with ethyl acetate. The extract waswashed with a saturated aqueous solution of sodium bicarbonate, waterand an aqueous solution of sodium chloride in turn, dried and thenevaporated to dryness under reduced pressure. The residue was stirredfor 30 minutes in diethyl ether, and the remained substance wascollected by filtration to give7-[2-(2-formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-methoxymethyl-3-cephem-4-carboxylicacid (syn isomer) (0.56 g).

IR (Nujol): 3250, 1780, 1660 cm⁻¹

NMR δppm (DMSO-d₆): 3.22 (3H, s), 3.55 (2H, broad s), 3.90 (3H, s), 4.19(2H, s), 5.17 (1H, d, J=5Hz), 5.81 (1H, dd, J=5Hz, 8Hz), 7.43 (1H, s),8.51 (1H, s), 9.67 (1H, d, J=8Hz), 12.58 (1H, broad s)

EXAMPLE 20

7-[2-(2-Formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid (syn isomer) (0.64 g) was obtained by reacting tert-butyl7-[2-(2-formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-methylthiomethyl-3-cephem-4-carboxylate(syn isomer) (0.9 g) with trifluoroacetic acid (5.8 g) in the presenceof anisole (0.9 ml) according to the similar manner to that of Example19.

IR (Nujol): 3250, 1780, 1660 cm⁻¹

NMR δppm (DMSO-d₆): 1.99 (3H, s), 3.38-4.1 (4H, m), 3.93 (3H, s), 5.25(1H, d, J=5Hz), 5.79 (1H, dd, J=5Hz, 8Hz), 7.45 (1H, s), 8.43 (1H, s),9.69 (1H, d, J=8Hz), 12.68 (1H, broad s)

The compounds described in the following Examples 21 and 22 wereobtained by reacting tert-butyl ester of the correspondingcephalosporanic acid derivatives with trifluoroacetic acid in thepresence of anisole according to the similar manner to that of Example19.

EXAMPLE 21

7-[2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-methoxymethyl-3-cephem-4-carboxylicacid hydrochloride (syn isomer)

IR (Nujol): 3300, 1780, 1720, 1660, 1640 cm⁻¹

EXAMPLE 22

7-[2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid (syn isomer)

IR (Nujol): 3350, 1770, 1670 cm⁻¹

EXAMPLE 23

To a suspension of7-[2-(2-formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-methoxymethyl-3-cephem-4-carboxylicacid (syn isomer) (0.52 g) in a mixture of methanol (3 ml) andtetrahydrofuran (2 ml) was added conc. hydrochloric acid (0.18 g), andthe mixture was stirred at 30° C. for 4 hours. After the reactionmixture was cooled and diluted with diisopropyl ether, the precipitatedcrystals were collected by filtration, washed with diisopropyl ether andthen dried to give7-[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-methoxymethyl-3-cephem-4-carboxylicacid hydrochloride (syn isomer) (0.45 g).

IR (Nujol): 3300, 1780, 1720, 1660, 1640 cm⁻¹

NMR δppm (DMSO-d₆): 3.26 (3H, s), 3.58 (2H, broad s), 4.0 (3H, s), 4.24(2H, s), 5.24 (1H, d, J=5Hz), 5.82 (1H, dd, J=5Hz, 8Hz), 7.01 (1H, s),9.87 (1H, d, J=8Hz)

EXAMPLE 24

7-[2-(2-Aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid (syn isomer) (0.23 g) was obtained by reacting7-[2-(2-formamidothiazol-4-yl)-2-methoxyiminoacetamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid (syn isomer) (0.6 g) with conc. hydrochloric acid (0.4 g) in amixture of methanol (3 ml) and tetrahydrofuran (1 ml) according to thesimilar manner to that of Example 23.

IR (Nujol): 3350, 1770, 1670 cm⁻¹

NMR δppm (DMSO-d₆): 2.01 (3H, s), 3.2-4.1 (4H, m), 3.88 (3H, s), 5.25(1H, d, J=5Hz), 5.78 (1H, dd, J=5Hz, 8Hz), 6.82 (1H, s), 7.24 (2H, broads), 9.64 (1H, d, J=8Hz)

EXAMPLE 25

To a solution of benzhydryl7-amino-3-methylthiomethyl-3-cephem-4-carboxylate (10 g) andtrimethylsilylacetamide (18.4 g) in methylene chloride (100 ml) wasadded at -15° C. an activated acid, which was prepared from(2-formamidothiazol-4-yl)glyoxylic acid (6.56 g), N,N-dimethylformamide(2.92 ml) and phosphorus oxychloride (3.46 ml) in a conventional manner,and the mixture was stirred at -20 ° to -15° C. for 15 minutes. Afterthe reaction mixture was poured into water, it was extracted with ethylacetate. The extract was washed with an aqueous solution of sodiumbicarbonate and an aqueous solution of sodium chloride, followed bydrying over anhydrous magnesium sulfate. Removal of the solvent gave aresidue (13.4 g), which was chromatographed on silica gel (100 ml) usinga mixture of benzene and ethyl acetate (5:1 by volume) as an eluent. Thefractions containing the desired compound were collected and thenevaporated to dryness to obtain benzhydryl7-[(2-formamidothiazol-4-yl)glyoxylamido]-3-methylthiomethyl-3-cephem-4-carboxylate (7.1 g).

I.R. (Nujol): 3300, 1780, 1700, 1656 cm⁻¹

N.M.R. δppm (DMSO-d₆): 1.83 (3H, s), 3.58 (2H, broad s), 3.67 (2H, broads), 5.33 (1H, d, J=5Hz), 5.87 (1H, dd, J=5Hz, 8Hz), 6.90 (1H, s), 7.40(10H, s), 8.47 (1H, s), 8.58 (1H, s), 9.88 (1H, d, J=8Hz), 12.68 (1H,broad s)

The following compounds were obtained by reacting 7-amino-3-substitutedcephalosporanic acid derivatives with the corresponding acids accordingto the similar manner to that of Example 25.

EXAMPLE 26

7-[(2-Aminothiazol-4-yl)glyoxylamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid.

I.R. (Nujol): 3300, 1762, 1522 cm⁻¹

EXAMPLE 27

7-[2-(2-Aminothiazol-4-yl)-DL-glycinamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid.

I.R. (Nujol): 3300, 1755, 1686, 1600 cm⁻¹

EXAMPLE 28

To a mixture of benzhydryl7-amino-3-methylthiomethyl-3-cephem-4-carboxylate (4.92 g) andN-tert-butoxycarbonyl-2-(2-formamidothiazol-4-yl)-DL-glycine (4.0 g) inmethylene chloride (100 ml) and tetrahydrofuran (80 ml) was addedN,N'-dicyclohexylcarbodiimide (2.39 g), and the mixture was stirred atambient temperature for an hour. After the insoluble substance wasremoved by filtration, the filtrate was evaporated to dryness to give aresidue, which was dissolved in ethyl acetate. This solution was washedwith an aqueous solution of sodium bicarbonate and an aqueous solutionof sodium chloride, and then dried over anhydrous magnesium sulfate.Removal of the solvent gave benzhydryl

7-[N-tert-butoxycarbonyl-2-(2-formamidothiazol-4-yl)-DL-glycinamido]-3-methylthiomethyl-3-cephem-4-carboxylate(9.4 g).

I.R. (Nujol): 3300, 1770, 1710, 1680, 1615 cm⁻¹

N.M.R. δppm (DMSO-d₆): 1.36 (9H, s), 1.80 (3H, s), 3.58 (4H, m),

    ______________________________________                                                 5.16 (d, J = 5 Hz)                                                                            (1H)                                                         5.23 (d, J = 5 Hz)                                                    ______________________________________                                    

5.38 (1H, d, J=8Hz), 5.6-5.9 (1H, m), 6.90 (1H, s), 7.13 (1H, s), 7.33(10H, broad s), 8.47 (1H, s), 9.08 (1H, d, J=8Hz)

EXAMPLE 29

A mixture of7-[(2-formamidothiazol-4-yl)-glyoxylamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid (3.0 g) and conc. hydrochloric acid (3 ml) in methanol (50 ml) wasstirred at ambient temperature for 2 hours. The reaction mixture wasadjusted to pH 5-6 with an aqueous solution of sodium bicarbonate andthen concentrated under reduced pressure. The precipitated crystals inthe concentrate were collected by filtration to give7-[(2-aminothiazol-4-yl)glyoxylamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid (1.1 g).

I.R. (Nujol): 3300, 1762, 1522 cm⁻¹

N.M.R. δppm (DMSO-d₆): 2.00 (3H, s), 3.65 (4H, broad s), 5.22 (1H, d,J=5Hz), 5.68 (1H, dd, J=5Hz, 8Hz), 7.37 (2H, broad s), 7.83 (1H, s),9.73 (1H, d, J=8Hz)

EXAMPLE 30

A mixture of benzhydryl7-[N-tert-butoxycarbonyl-2-(2-formamidothiazol-4-yl)-DL-glycinamido]-3-methylthiomethyl-3-cephem-4-carboxylate(9.4 g) and conc. hydrochloric acid (4.16 ml) in methanol (100 ml) andtetrahydrofuran (25 ml) was stirred at 30° to 35° C. for 5 hours. Thereaction mixture was adjusted to pH 4.5 with sodium bicarbonate and thenevaporated to dryness to give a residue, which was dissolved in ethylacetate. This solution was washed with an aqueous solution of sodiumchloride, dried over anhydrous magnesium sulfate and then evaporated toobtain benzhydryl7-[N-tert-butoxycarbonyl-2-(2-aminothiazol-4-yl)-DL-glycinamido]-3-methylthiomethyl-3-cephem-4-carboxylate(7.4 g).

I.R. (Nujol): 3300, 1772, 1716, 1685, 1623, 1244, 1170, 1160 cm⁻¹

N.M.R. δppm (DMSO-d₆): 1.40 (9H, s), 1.80 (3H, s), 3.6 (4H, m), 5.0-5.4(2H, m), 5.6-5.9 (2H, m), 6.93 (1H, s), 7.30 (1H, s), 7.43 (10H, broads), 8.93 (1H, d, J=8Hz)

The following compounds were obtained by reacting7-acylamino-3-substituted cephalosporanic acid derivatives having aformamido group with hydrochloric acid according to the similar mannerto that of Example 30.

EXAMPLE 31

7-[2-(2-Aminothiazol-4-yl)-DL-glycinamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid.

I.R. (Nujol): 3300, 1755, 1686, 1600 cm⁻¹

EXAMPLE 32

7-[2-(2-Aminothiazol-4-yl)-DL-glycolamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid.

I.R. (Nujol): 3300, 1752, 1675, 1600 cm⁻¹

EXAMPLE 33

Benzhydryl7-[(2-formamidothiazol-4-yl)glyoxylamido]-3-methylthiomethyl-3-cephem-4-carboxylate(7.0 g) was dissolved in a solution of methylene chloride (70 ml),anisole (7 ml) and trifluoroacetic acid (14 ml), and the mixture wasstirred at ambient temperature for an hour. After the solvent wasremoved by distillation under reduced pressure, the residue wasdissolved in water, adjusted to pH 2.0 with conc. hydrochloric acid andthen extracted with ethyl acetate. The extract was washed with anaqueous sodium chloride and dried over magnesium sulfate, followed byevaporation. The residue was pulverized with diisopropyl ether to obtain7-[(2-formamidothiazol-4-yl)glyoxylamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid (3.3 g).

I.R. (Nujol): 3120, 1762, 1731, 1677 cm⁻¹

N.M.R. δppm (DMSO-d₆): 2.00 (3H, s), 3.63 (4H, broad s), 5.24 (1H, d,J=5Hz), 5.73 (1H, dd, J=5Hz, 8Hz), 8.43 (1H, s), 8.57 (1H, s), 9.90 (1H,d, J=8Hz), 12.80 (1H, broad s)

The following compounds were obtained by reacting7-acylamino-3-substituted cephalosporanic acid derivatives havingbenzhydryl ester with 2,2,2-trifluoroacetic acid in the presence ofanisole according to the similar manner to that of Example 33.

EXAMPLE 34

7-[(2-Aminothiazol-4-yl)glyoxylamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid.

I.R. (Nujol): 3300, 1762, 1522 cm⁻¹

EXAMPLE 35

7-[2-(2-Aminothiazol-4-yl)-DL-glycolamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid.

I.R. (Nujol): 3300, 1752, 1675, 1600 cm⁻¹

EXAMPLE 36

A mixture of benzhydryl7-[N-tert-butoxycarbonyl-2-(2-aminothiazol-4-yl)-DL-glycinamido]-3-methylthiomethyl-3-cephem-4-carboxylate(7.0 g), anisole (7 ml) and 2,2,2-trifluoroacetic acid (21 ml) wasstirred at 5° C. for half an hour. To the reaction mixture was addeddropwise diisopropyl ether (300 ml), and the precipitated substance wascollected by filtration and then washed with diisopropyl ether, followedby dissolving in a mixture of ethyl acetate (50 ml) and water (100 ml).After the aqueous layer was separated out, the ethyl acetate in theaqueous layer was completely removed, and then the remained aqueoussolution was adjusted to pH 4.2 with 5% aqueous solution of sodiumbicarbonate. The resultant aqueous solution was chromatographed onnonionic adsorption resin "Diaion HP-20" (Trade Mark, made by MitsubishiChemical Industries Ltd.) (100 ml). After washing with water (250 ml),elution was carried out with 30% aqueous isopropyl alcohol. Thefractions containing the desired compound were collected and thenevaporated, followed by lyophilization to obtain7-[2-(2-aminothiazol-4-yl)-DL-glycinamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid (2.9 g).

I.R. (Nujol): 3300, 1755, 1686, 1600 cm⁻¹

N.M.R. δppm (D₂ O+DCl): 2.07 (3H, s), 3.5-4.0 (4H, m), 5.27 (1H, d,J=5Hz), 5.50 (1H, s),

    ______________________________________                                                 5.60 (d, J = 5Hz)                                                                             (1H),                                                        5.72 (d, J = 5Hz)                                                     ______________________________________                                    

7.27 (1H, s)

EXAMPLE 37

To a solution of7-[(2-aminothiazol-4-yl)glyoxylamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid (1.1 g) in methanol (80 ml) was added sodium borohydride (150 mg)at 5° to 10° C., and the mixture was stirred at the same temperature forhalf an hour. After the reaction mixture was adjusted to pH 5.0 with 10%hydrochloric acid, the solvent was removed by distillation. To theresidue was added water (50 ml), followed by adjusting to pH 5.0 with10% hydrochloric acid. The resultant aqueous solution waschromatographed on nonionic adsorption resin "Diaion HP-20" (50 ml).After washing with water (100 ml), elution was carried out with 30%aqueous isopropyl alcohol. The fractions containing the desired compoundwere collected and then evaporated to dryness, followed bylyophilization to obtain7-[2-(2-aminothiazol-4-yl)-DL-glycolamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid (0.75 g).

I.R. (Nujol): 3300, 1752, 1675, 1600 cm⁻¹

N.M.R. δppm (DMSO-d₆): 2.00 (3H, s), 3.48 (2H, broad s), 3.67 (2H, broads), 4.93 (1H, s), 5.07 (1H, d, J=5Hz), 5.57 (1H, m), 6.50 (1H, s), 7.03(2H, broad s),

    ______________________________________                                                 8.33 (d, J = 8 Hz)                                                                            (1H)                                                         8.42 (d, J = 8 Hz)                                                    ______________________________________                                    

EXAMPLE 38

Vilsmeier reagent prepared from phosphorus oxychloride (1.23 ml) andN,N-dimethylformamide (1.1 ml) was suspended in dry tetrahydrofuran (40ml). To the suspension was added2-(2-formamidothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetic acid(syn isomer) (4.0 g) under ice-cooling with stirring, and the mixturewas stirred at the same temperature for 50 minutes to prepare theactivated acid solution. On the other hand, benzhydryl7-amino-3-methylthiomethyl-3-cephem-4-carboxylate (4.66 g) andtrimethylsilylacetamide (8.6 g) were dissolved in methylene chloride (50ml). To the solution was added the activated acid solution at -20° C. ata time, and the mixture was stirred at the same temperature for an hour.Water (100 ml) and ethyl acetate (200 ml) were added to the resultantsolution, and the organic layer was separated, washed with 5% aqueoussodium bicarbonate and an aqueous sodium chloride, and then dried overmagnesium sulfate, followed by evaporation under reduced pressure togive benzhydryl7-[2-(2-formamidothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetamido]-3-methylthiomethyl-3-cephem-4-carboxylate (syn isomer) (8.0 g), mp132°-138° C.

IR (Nujol): 3260, 1783, 1725, 1687 cm⁻¹

NMR δppm (DMSO-d6δ): 1.47 (9H, s), 1.83 (3H, s), 3.60 (2H, broad s),3.66 (2H, broad s), 4.98 (2H, broad s), 5.32 (1H, d, J=5Hz), 5.92 (1H,dd, J=5Hz, 8Hz), 6.95 (1H, s), 7.4 (10H, m), 7.48 (1H, s), 8.54 (1H, s),9.63 (1H, d, J=8Hz), 12.65 (1H, broad s)

EXAMPLE 39

Benzhydryl7-[2-(2-formamidothiazol-4-yl)-2-benzhydryloxycarbonylmethoxyiminoacetamido]-3-methylthiomethyl-3-cephem-4-carboxylate(syn isomer) (7.8 g) was obtained by reacting benzhydryl7-amino-3-methylthiomethyl-3-cephem-4-carboxylate (4.0 g) with theactivated acid solution prepared from2-(2-formamidothiazol-4-yl)-2-benzhydryloxycarbonylmethoxyiminoaceticacid (syn isomer) (6.18 g), phosphorus oxychloride (1.42 ml) andN,N-dimethylformamide (1.21 ml), according to a similar manner to thatof Example 38, mp 135°-142° C.

IR (Nujol): 3250, 1780, 1722, 1685 cm⁻¹

NMR δppm (DMSO-d6): 1.83 (3H, s), 3.63 (4H, m), 5.0 (2H, broad s), 5.35(1H, d, J=5Hz), 5.98 (1H, dd, J=5Hz, 8Hz), 6.95 (1H, s), 6.98 (1H, s),7.37 (20H, m), 7.50 (1H, s), 8.57 (1H, s), 9.82 (1H, d, J=8Hz), 12.73(1H, broad s)

EXAMPLE 40

Benzhydryl7-[2-benzhydryloxycarbonylmethoximino-2-{2-(2,2,2-trifluoroacetamido)thiazol-4-yl}acetamido]-3-methylthiomethyl-3-cephem-4-carboxylate(syn isomer) (5.6 g) was obtained by reacting benzhydryl7-amino-3-methylthiomethyl-3-cephem-4-carboxylate (3.0 g) with theactivated acid solution prepared from2-benzhydryloxycarbonylmethoximino-2-[2-(2,2,2-trifluoroacetamido)thiazol-4-yl]aceticacid (syn isomer) (3.68 g), phosphorus oxychloride (0.89 ml) andN,N-dimethylformamide (0.75 ml), according to a similar manner to thatof Example 38, mp 165°-169° C.

IR (Nujol): 3300, 1786, 1733, 1675, 1610 cm⁻¹

NMR δppm (DMSO-d6): 2.00 (3H, s), 3.56 (4H, m), 4.84 (2H, s), 5.26 (1H,d, J=5Hz), 5.86 (1H, dd, J=5Hz, 8Hz), 6.84 (1H, s), 6.88 (1H, s), 7.3(20H, m), 9.62 (1H), d, J=8Hz)

EXAMPLE 41

To a suspension of benzhydryl7-[2-(2-formamidothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetamido]-3-methylthiomethyl-3-cephem-4-carboxylate(syn isomer) (8.0 g) in methanol (160 ml) was added conc. hydrochloricacid (5.6 ml), and the mixture was stirred at 35° C. for an hour. Theresultant solution was adjusted to pH 5.0 with a saturated aqueoussodium bicarbonate. After distilling methanol under reduced pressure,the residue was dissolved in water (100 ml) and ethyl acetate (200 ml).The ethyl acetate layer was washed with a saturated aqueous sodiumchloride and dried over magnesium sulfate. The solvent was removed byfiltration to give benzhydryl7-[2-(2-aminothiazol-4-yl)-2-tert-butoxycarbonylmethoxyiminoacetamido]-3-methylthiomethyl-3-cephem-4-carboxylate(syn isomer) (7.0 g), mp 140°-145° C.

IR (Nujol): 3250, 1780, 1723, 1680 cm⁻¹

NMR δppm (DMSO-d6): 1.43 (9H, s), 1.83 (3H, s), 3.63 (4H, m), 4.6 (2H,broad s), 5.29 (1H, d, J=5Hz), 5.87 (1H, dd, J=5Hz, 8Hz), 6.83 (1H, s),6.95 (1H, s), 7.4 (10H, m), 9.52 (1H, d, J=8Hz)

EXAMPLE 42

Benzhydryl7-[2-(2-aminothiazol-4-yl)-2-benzhydryloxycarbonylmethoxyiminoacetamido]-3-methylthiomethyl-3-cephem-4-carboxylate(syn isomer) (7.0 g), mp 148°-155° C., was obtained by reactingbenzhydryl7-[2-(2-formamidothiazol-4-yl)-2-benzhydryloxycarbonylmethoxyiminoacetamido]-3-methylthiomethyl-3-cephem-4-carboxylate(syn isomer) (7.5 g) with conc. hydrochloric acid (3.9 ml) according toa similar manner to that of Example 41.

NMR δppm (DMSO-d6): 1.83 (3H, s), 3.60 (4H, m), 4.90 (2H, broad s), 5.30(1H, d, J=5Hz), 5.87 (1H, dd, J=5Hz, 8Hz), 6.86 (1H, s), 6.93 (1H, s),6.97 (1H, s), 7.4 (20H, m), 9.67 (1H, d, J=8Hz).

EXAMPLE 43

A solution of 7-[2-carboxymethoxyimino-2-{2-(2,2,2-trifluoroacetamido)thiazol-4-yl}acetamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid (syn isomer) (4.1 g) and sodium acetate (9.56 g) in water (41 ml)was stirred for 19.5 hours at ambient temperature. The resultantsolution was adjusted to pH 2.0 with conc. hydrochloric acid. Theprecipitates were collected by filtration and dried over phosphoruspentoxide to give7-[2-carboxymethoxyimino-2-(2-aminothiazol-4-yl)acetamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid (syn isomer) (1.8 g), mp 173°-176° C. (dec.).

IR (Nujol): 3370, 1772, 1670 (broad) cm⁻¹

NMR δppm (DMSO-d6): 2.00 (3H, s), 3.57 (4H, m), 4.60 (2H, broad s), 5.17(1H, d, J=5Hz), 5.73 (1H, dd, J=5Hz, 8Hz), 6.77 (1H, s), 9.45 (1H, d,J=8Hz)

EXAMPLE 44

To a solution of benzhydryl7-[2-benzhydryloxycarbonylmethoxyimino-2-{2-(2,2,2-trifluoroacetamido)-thiazol-4-yl}acetamido]-3-methylthiomethyl-3-cephem-4-carboxylate(syn isomer) (5.6 g) and anisole (5.6 ml) in methylene chloride (11.2ml) was added trifluoroacetic acid (11.2 ml) at 10° C. The mixture wasstirred for 1.5 hours at ambient temperature and then poured into amixture of diisopropyl ether (400 ml) and petroleum ether (100 ml). Theprecipitates were collected by filtration and washed with petroleumether to give7-[2-carboxymethoxyimino-2-{2-(2,2,2-trifluoroacetamido)-thiazol-4-yl}acetamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid (syn isomer) (4.2 g), mp 183°-186° C. (dec.).

IR (Nujol): 3300, 1778, 1723, 1660 cm⁻¹

NMR δppm (DMSO-d6): 2.00 (3H, s), 3.62 (4H, m), 4.70 (2H, s), 5.24 (1H,d, J=5Hz), 5.80 (1H, dd, J=5Hz, 8Hz), 7.60 (1H, s), 9.68 (1H, d, J=8Hz)

What we claim is:
 1. A compound which is7[2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]-3-methylthiomethyl-3-cephem-4-carboxylicacid or carboxylic ester thereof, or a pharmaceutically acceptable saltthereof.
 2. An antimicrobial pharmaceutical composition comprising, asan antimicrobially active ingredient, an effective amount of a compoundof claim 1, in admixture with a pharmaceutically acceptable carrier. 3.A method for treating an infectious disease caused by pathogenicmicroorganisms which comprises administering an effective amount of acompound of claim 1 to infected human beings or animals.